BACKGROUND As living biodrugs,mesenchymal stem cells(MSCs)have progressed to phase 3 clinical trials for cardiovascular applications.However,their limited immediate availability hampers their routine clinical use.AIM ...BACKGROUND As living biodrugs,mesenchymal stem cells(MSCs)have progressed to phase 3 clinical trials for cardiovascular applications.However,their limited immediate availability hampers their routine clinical use.AIM To validate our hypothesis that cryopreserved MSCs(CryoMSCs)are as safe and effective as freshly cultured MSC counterparts but carry logistical advantages.METHODS Four databases were systematically reviewed for relevant randomized controlled trials(RCTs)evaluating the safety and efficacy of CryoMSCs from various tissue sources in treating patients with heart disease.A subgroup analysis was performed based on MSC source and post-thaw cell viability to determine treatment effects across different CryoMSCs sources and viability status.Weighted mean differences(WMDs)and odds ratios were calculated to measure changes in the estimated treatment effects.All statistical analyses were performed using RevMan version 5.4.1 software.RESULTS Seven RCTs(285 patients)met the eligibility criteria for inclusion in the metaanalysis.During short-term follow-up,^(Cryo)MSCs demonstrated a significant 2.11%improvement in left ventricular ejection fraction(LVEF)[WMD(95%CI)=2.11(0.66-3.56),P=0.004,I2=1%],with umbilical cord-derived MSCs being the most effective cell type.However,the significant effect on LVEF was not sustained over the 12 months of follow-up.Subgroup analysis demonstrated a substantial 3.44%improvement in LVEF[WMD(95%CI)=3.44(1.46-5.43),P=0.0007,I2=0%]when using MSCs with post-thaw viability exceeding 80%.There was no statistically significant difference in the frequency of major cardiac adverse events observed in rehospitalization or mortality in patients treated with ^(Cryo)MSCs vs the control group.CONCLUSION ^(Cryo)MSCs are a promising option for heart failure patients,particularly considering the current treatment options for cardiovascular diseases.Our data suggest that ^(Cryo)MSCs could be a viable alternative or complementary treatment to the current options,potentially improving patient outcomes.展开更多
To Editor,We read with interest the article entitled ＂Transplantation of cryopreserved teeth： a systematic review＂ （Osathanon, 2010）. Although the author is to be congratulated for his systematic approach to cryo...To Editor,We read with interest the article entitled ＂Transplantation of cryopreserved teeth： a systematic review＂ （Osathanon, 2010）. Although the author is to be congratulated for his systematic approach to cryopreserved tooth transplantation （CTT）, we would like to draw your attention to some technical limitations of this review.展开更多
The ocular surface is covered by an epithelium encompassing an area including the cornea,the limbus and the conjunctiva bordered by the upper and lower lids.The healthy state of the ocular surface epithelium depends o...The ocular surface is covered by an epithelium encompassing an area including the cornea,the limbus and the conjunctiva bordered by the upper and lower lids.The healthy state of the ocular surface epithelium depends on a stable and protective preocular tear film when the eye is open.A stable preocular tear film is governed by sound ocular surface defense that involves effective展开更多
Objective: This study reports the outcomes of the cryopreserved mitral homograft in 119 patients prospectively followed with clinical, echocardiographic and structural valve deterioration assessments. Methods: 119 pat...Objective: This study reports the outcomes of the cryopreserved mitral homograft in 119 patients prospectively followed with clinical, echocardiographic and structural valve deterioration assessments. Methods: 119 patients undergoing mitral and aortic homograft implantation. Patient’s causes of mitral disease were: rheumatic disease (n = 75), endocarditis (n = 37) and others (n = 7). There were 40 partial homografts and 88 total homografts. Mitro-aortic homograft valve replacement was performed in 29 cases. Results: Mean follow-up was 9.8 ± 6.3 years (up to 19.2 yrs). There were 7 early (2展开更多
Aim: To 1) compare post-wash and post-thaw parameters of sperm processed with PureSperm density gradient technique and swim-up method; and 2) test the efficacy of two commonly available density gradient media PureSper...Aim: To 1) compare post-wash and post-thaw parameters of sperm processed with PureSperm density gradient technique and swim-up method; and 2) test the efficacy of two commonly available density gradient media PureSperm and ISolate. Methods: This prospective study used semen specimens from 22 patients. Specimens from nine patients were processed by both PureSperm density gradient and swim-up method. These specimens were then cryopreserved. Thirteen specimens were processed by both PureSperm (40 % and 80 %) and Isolate (50 % and 90 %) double density gradient techniques. The two fractions processed by both PureSperm and swim-up were analyzed for post-wash sperm characteristics. Post-thaw analysis was done after 24 hours. Sperm fractions obtained after processing with PureSperm and ISolate were compared for post-wash sperm characteristics and ROS levels. Results: Specimens prepared with PureSperm had significantly higher median total motile sperm counts (TMSC) (32.2 x 10~6 vs. 17.6 x 10~6), recovery rates (69.2 % vs. 50.0 %), and longevity at 4 hours (83.0 % vs. 55.0 %) compared to specimen prepared by swim-up. Post-thaw specimens also had a higher recovery and longevity at 4 hours with PureSperm as compared to the swim-up. Semen specimens processed by PureSperm had significantly higher total sperm count, TMSC, and percentage recovery rates (30.0 % vs. 19.7 %) than ISolate. Conclusion: Semen quality is better preserved in fresh and cryopreserved semen prepared with PureSperm density gradient compared to swim-up. A significant enrichment of sperm is observed with PureSperm compared to ISolate. Higher recovery rates of mature motile sperm obtained after PureSperm sperm preparation may be beneficial for successful ART. i展开更多
This study is aimed at investigating the potentials of ex vivo expansion and pluri-differentiation of cryopreservation of adult human bone marrow mesenchymal stem cells （hMSCs） into chondrocytes, adipocytes and neur...This study is aimed at investigating the potentials of ex vivo expansion and pluri-differentiation of cryopreservation of adult human bone marrow mesenchymal stem cells （hMSCs） into chondrocytes, adipocytes and neurocytes. Cryopreserved hMSCs were resuscitated and cultured for 15 passages, and then induced into chondrocytes, adipocytes and neurocytes with corresponding induction medium. The induced cells were observed for morphological properties and detected for expressions of type II collagen, triglyceride or neuron-specific enolase and nestin. The result showed that the resuscitated cells could differentiate into chondrocytes after exposure to transforming growth factor 61 （TGF-~0, insulin-like growth factor I （IGF-I） and vitamin C （Vc）, and uniformly changed morphologically from a spindle-like fibroblastic appearance to a polygonal shape in three weeks. The induced cells were heterochromatic to safranin O and expressed cartilage matrix-procollagenal （If） mRNA. The resuscitated cells cultured in induction medium consisting of dexamethasone, 3-isobutyl-l-methylxanthine, indomethacin and IGF-I showed adipogenesis, and lipid vacuoles accumulation was detectable after 21 d. The resuscitated hMSCs were also induced into neurocytes and expressed nestin and neuron specific endolase （NSE） that were special surface markers associated with neural cells at different stage. This study suggested that the resuscitated hMSCs should be still a population ofpluripotential cells and that it could be used for establishing an abundant bMSC reservoir for further experiment and treatment of various clinical discases.展开更多
Cryopreservation is associated with the production of reactive oxygen species, which leads to lipid peroxidation of the sperm membrane and consequently a reduction in sperm motility and decreased fertility potential. ...Cryopreservation is associated with the production of reactive oxygen species, which leads to lipid peroxidation of the sperm membrane and consequently a reduction in sperm motility and decreased fertility potential. The aim of this study was to determine the optimal concentration of L-cysteine needed for cryopreservation of boar semen. Twelve boars provided semen of proven motility and morphology for this study. The semen was divided into four portions in which the lactose-egg yolk （LEY） extender used to resuspend the centrifuged sperm pellet was supplemented with various concentrations of L-cysteine to reach 0 mmol L^-1 （group Ⅰ, control）, 5 mmol L^-1 （group Ⅱ）, 10 mmol L^-1 （group Ⅲ） and 15 mmol L^-1 （group Ⅳ）. Semen suspensions were loaded in straws （0.5 mL） and placed in a controlled-rate freezer. After cryopreservation, frozen semen samples were thawed and investigated for progressive motility, viability using SYBR-14/EthD-1 staining and acrosome integrity using FITC-PNA/EthD-1 staining. There was a significantly higher （P 〈 0.01） percentage of progressive motility, viability and acrosomal integrity in two L-cysteine-supplemented groups （group Ⅱ and group Ⅲ） compared with the control. There was a biphasic effect of L-cysteine, with the highest percentage of progressive motility, viability and acrosomal integrity in group Ⅲ. In conclusion, 5 or 10 mmol L^-1 was the optimum concentration of L-cysteine to be added to the LEY extender for improving the quality of frozen-thawed boar semen.展开更多
We performed this study to evaluate the clinical outcomes of microdissection testicular sperm extraction-intracytoplasmic sperm injection(micro-TESE-ICSI)treatment that used fresh or cryopreserved sperm in patients wi...We performed this study to evaluate the clinical outcomes of microdissection testicular sperm extraction-intracytoplasmic sperm injection(micro-TESE-ICSI)treatment that used fresh or cryopreserved sperm in patients with nonobstructive azoospermia(NOA).A total of 338 NOA patients with 344 consecutive cycles received treatment in the reproductive medicine center of Peking University Third Hospital in Beijing,China,from January 2014 to December 2017.Fresh oocytes and fresh sperm were used in 222 patients with 234 cycles(Group A).Fresh oocytes and cryopreserved sperm were used in 116 patients with 110 cycles(Group B).We compared patient characteristics,embryonic development,and pregnancy outcomes between Groups A and B.There was no statistical difference in the patient characteristics,and no differences were observed with fertilization or quality embryo rates between Groups A and B.The rates of clinical pregnancy and live birth were both higher for Group A than those for Group B(both P<0.05).In conclusion,fresh testicular sperm appears to produce better ICSI outcomes than cryopreserved testicular sperm in patients with NOA.展开更多
The aim of this article was to examine the research articles regarding biological and mechanical properties of cryopreserved teeth for potential use in tooth transplantation. A systematic review of literatures was per...The aim of this article was to examine the research articles regarding biological and mechanical properties of cryopreserved teeth for potential use in tooth transplantation. A systematic review of literatures was performed by Pubmed searching with assigned key words from January 1, 1990 to June 8, 2009. All articles were examined for inclusion criteria. Secondary search was conducted by hand-search through references of included articles from primary search. A total of 24 articles were obtained from both primary and secondary search and used as fundamental articles in this review. Periodontal ligament tissues of cryopreserved teeth were able to maintain their biological properties resulted in a satisfactory healing of periodontium. Dental pulp tissues,however, may be compromised by limitation of permeability of cryopreservative agent into pulp cavity. Therefore, an endodontic treatment of transplanted cryopreserved teeth was recommended. Cryopreserved teeth had comparable mechanical properties to those of normal teeth. Importantly, the success of cryopreserved tooth transplantation treatment in orthodontic patients was reported. The cryopreserved teeth for tooth banking have a potential clinical application for treatment of missing teeth. Case selection, however, is critical for treatment success. More studies and data regarding masticatory function and periodontal healing of transplanted cryopreserved teeth are needed.展开更多
Objective： To evaluate the clinical efficacy of peripheral deep anterior lamellar keratoplasty （DALK） using a cryopreserved donor cornea for Terrien＇s marginal degeneration （TMD）. Methods： Thirty-one eyes of 27...Objective： To evaluate the clinical efficacy of peripheral deep anterior lamellar keratoplasty （DALK） using a cryopreserved donor cornea for Terrien＇s marginal degeneration （TMD）. Methods： Thirty-one eyes of 27 patients with TMD underwent peripheral DALK using cryopreserved donor corneas, According to the distance between the inner edge of the lesion and the limbus, a ring-shaped or D-shaped DALK was performed. All grafts were stored at -20 ℃. Cryopreserved comeoscleral rims were prepared for ring-shaped grafts and cryopreserved whole eyeballs were prepared for D-shaped grafts. The general conditions, intraoperative performance, postoperative corneal reconstruc- tion, astigmatism, best corrected visual acuity （BCVA）, and various complications were analyzed. Results： Ring-shaped DALK was performed in 28 eyes and D-shaped DALK was performed in 3 eyes. Postoperative follow-up time was （28.4±24.8） months. There was evidence of inflammation before surgery in 12 eyes （38.7%） and intraoper- ative perforation occurred in 13 eyes （41.9%）. The corneal structures of all eyes were reconstructed. Postoperative astigmatism and BCVA showed improvement （both P=0.00） except for cases that underwent D-shaped DALK. Ten eyes （32.2%） developed transient ocular hypertension and one eye （3.2%） developed secondary glaucoma. No pri- mary disease recurrence or corneal allograft rejection was observed. Conclusions： Peripheral DALK for TMD using cryopreserved donor tissue is an effective technique that eliminates rejection and extends the use of donor eyes. Inflammatory history or intraoperative perforation has no adverse effect on graft recovery. However, D-shaped DALK did not achieve good visual outcomes.展开更多
The experimental design evaluated histological,mechanical,and biological properties of allogeneic decellularized nerves after cryopreservation in a multi-angle,multi-directional manner to provide evidence for long-ter...The experimental design evaluated histological,mechanical,and biological properties of allogeneic decellularized nerves after cryopreservation in a multi-angle,multi-directional manner to provide evidence for long-term preservation.Acellular nerve allografts from human and rats were cryopreserved in a cryoprotectant（10% fetal bovine serum,10% dimethyl sulfoxide,and 5% sucrose in RPMI1640 medium） at-80°C for 1 year,followed by thawing at 40°C or 37°C for 8 minutes.The breaking force of acellular nerve allografts was measured using a tensile test.Cell survival was determined using L-929 cell suspensions.Acellular nerve allografts were transplanted into a rat model with loss of a 15-mm segment of the left sciatic nerve.Immunohistochemistry staining was used to measure neurofilament 200 expression.Hematoxylin-eosin staining was utilized to detect relative muscle area in gastrocnemius muscle.Electron microscopy was applied to observe changes in allograft ultrastructure.There was no obvious change in morphological appearance or ultrastructure,breaking force,or cytotoxicity of human acellular nerve allografts after cryopreservation at-80°C.Moreover,there was no remarkable change in neurofilament 200 expression,myelin sheath thickness,or muscle atrophy when fresh or cryopreserved rat acellular nerve allografts were applied to repair nerve injury in rats.These results suggest that cryopreservation can greatly extend the storage duration of acellular nerve tissue allografts without concomitant alteration of the physiochemical and biological properties of the engineered tissue to be used for transplantation.展开更多
The relative merit of four different methods (method of fine tube, semen pellets, ampule, and test tube)of cryopreserved spermatozoa from 43 male donors with normal fertile ability was evaluated.The mobility and viabi...The relative merit of four different methods (method of fine tube, semen pellets, ampule, and test tube)of cryopreserved spermatozoa from 43 male donors with normal fertile ability was evaluated.The mobility and viability in post-thawed 172 samples were examined. The method of fine tube was recommedned to be the method of choice. A discussion on these four methods was given.展开更多
Objective： To study the method of cryopreserving rat hepatocytes and double collagen gel culture measurement after its cryopreservation. Methods： Rat hepatocytes, isolated by two-step perfusion with collagenase usin...Objective： To study the method of cryopreserving rat hepatocytes and double collagen gel culture measurement after its cryopreservation. Methods： Rat hepatocytes, isolated by two-step perfusion with collagenase using an extra corporeal perfusion apparatus, were cryopreserved in double collagen gel with culture medium added by epidermal growth factor（EGF）. The expression of cell function and cellular morphology were examined during culture. Results： The hepatocytes cryopreserved in double collagen gel concluding EGF showed good morphology and biological characteristics. After thawing, the MTT metabolism and protein synthesis of hepatocytes in sandwich ± EGF groups were better than those in control group. And the morphology and function of hepatocytes in sandwich group was better than that in EGF group（P 〈 0.05）. Conclusion： Double collagen gel culture can keep hepatocyte＇s activities. Thawed hepatocytes can be cultivated with collagenous matrix, which provides an environment that more closely resembles that in vivo and maintain the expression of certain liver-specific function of hepatocytes.展开更多
This study aimed to evaluate the outcomes and described the recovery process of cryopreserved limbal lamellar keratoplasty（CLLK） for peripheral corneal and limbal diseases. Thirteen eyes of 12 patients with a mean a...This study aimed to evaluate the outcomes and described the recovery process of cryopreserved limbal lamellar keratoplasty（CLLK） for peripheral corneal and limbal diseases. Thirteen eyes of 12 patients with a mean age of 41±23.9 y were included. The average follow-up was 12.1±5.6 mo. Stable ocular surface was achieved in all eyes at last follow-up. Epithelialization originated from both recipient and graft in 9 eyes. We conclude that CLLK compensates for the shortage of donor corneas and cryopreserved limbal grafts provide epithelialization sources in ocular surface reconstruction.展开更多
Objective:To test the effects of extenders and packaging methods on morphological and functional characteristics of frozen thawed Ossimi ram semen.Methods:Ram semen was pooled,diluted in 3 different extenders:Tris-egg...Objective:To test the effects of extenders and packaging methods on morphological and functional characteristics of frozen thawed Ossimi ram semen.Methods:Ram semen was pooled,diluted in 3 different extenders:Tris-egg yolk(TEY),Tris-soybean lecithin(TSBL),and Tris-butylatedhydroxytoluene(TBHT),equilibrated at 5℃for 4 h,and packaged in straws or pellets for freezing.Semen was evaluated for sperm progressive motility,viability,abnormality,and membrane integrity after dilution,equilibration and thawing.The percentages of viable,early apoptotic,apoptotic,and necrotic spermatozoa as well as comet assay parameters were determined in post-thawed semen.Total antioxidants capacity,malondialdehyde and lactic dehydrogenase were assayed in thawed seminal plasma.Results:After equilibration,only sperm membrane integrity was significantly higher(P<0.05)in TEY and TSBL than in TBHT.After thawing,TEY or TSBL in straws significantly improved sperm progressive motility and vitality(P<0.05).In thawed seminal plasma,TBHT or TSBL in straws and TBHT in pellets significantly reduced malondialdehyde,and TBHT in pellets significantly increased lactic dehydrogenase(P<0.05).TEY in straws increased viable sperm,while significantly decreased early apoptotic and apoptotic sperm(P<0.05).DNA damage was significantly decreased(P<0.05)in straws with TEY and TSBL,and tail moment decreased in straws with all extenders(P<0.05).Conclusions:Despite the disadvantages of dilution of cryopreserved semen with egg yolk,ram semen cryopreserved with TEY gives the best physical,morphological and functional characteristics in straws compared with pellets,followed by semen diluted with TSBL.However,semen diluted with TBHT or TSBL,regardless of packaging method,showed the highest impact on antioxidant status of cyopreserved ram semen.展开更多
文摘BACKGROUND As living biodrugs,mesenchymal stem cells(MSCs)have progressed to phase 3 clinical trials for cardiovascular applications.However,their limited immediate availability hampers their routine clinical use.AIM To validate our hypothesis that cryopreserved MSCs(CryoMSCs)are as safe and effective as freshly cultured MSC counterparts but carry logistical advantages.METHODS Four databases were systematically reviewed for relevant randomized controlled trials(RCTs)evaluating the safety and efficacy of CryoMSCs from various tissue sources in treating patients with heart disease.A subgroup analysis was performed based on MSC source and post-thaw cell viability to determine treatment effects across different CryoMSCs sources and viability status.Weighted mean differences(WMDs)and odds ratios were calculated to measure changes in the estimated treatment effects.All statistical analyses were performed using RevMan version 5.4.1 software.RESULTS Seven RCTs(285 patients)met the eligibility criteria for inclusion in the metaanalysis.During short-term follow-up,^(Cryo)MSCs demonstrated a significant 2.11%improvement in left ventricular ejection fraction(LVEF)[WMD(95%CI)=2.11(0.66-3.56),P=0.004,I2=1%],with umbilical cord-derived MSCs being the most effective cell type.However,the significant effect on LVEF was not sustained over the 12 months of follow-up.Subgroup analysis demonstrated a substantial 3.44%improvement in LVEF[WMD(95%CI)=3.44(1.46-5.43),P=0.0007,I2=0%]when using MSCs with post-thaw viability exceeding 80%.There was no statistically significant difference in the frequency of major cardiac adverse events observed in rehospitalization or mortality in patients treated with ^(Cryo)MSCs vs the control group.CONCLUSION ^(Cryo)MSCs are a promising option for heart failure patients,particularly considering the current treatment options for cardiovascular diseases.Our data suggest that ^(Cryo)MSCs could be a viable alternative or complementary treatment to the current options,potentially improving patient outcomes.
文摘To Editor,We read with interest the article entitled ＂Transplantation of cryopreserved teeth： a systematic review＂ （Osathanon, 2010）. Although the author is to be congratulated for his systematic approach to cryopreserved tooth transplantation （CTT）, we would like to draw your attention to some technical limitations of this review.
基金The development of PROKERA^(█)was supported in part with grant number EY014768 from the National Institute of Health(NIH)National Eye Institute(NEI)
文摘The ocular surface is covered by an epithelium encompassing an area including the cornea,the limbus and the conjunctiva bordered by the upper and lower lids.The healthy state of the ocular surface epithelium depends on a stable and protective preocular tear film when the eye is open.A stable preocular tear film is governed by sound ocular surface defense that involves effective
文摘Objective: This study reports the outcomes of the cryopreserved mitral homograft in 119 patients prospectively followed with clinical, echocardiographic and structural valve deterioration assessments. Methods: 119 patients undergoing mitral and aortic homograft implantation. Patient’s causes of mitral disease were: rheumatic disease (n = 75), endocarditis (n = 37) and others (n = 7). There were 40 partial homografts and 88 total homografts. Mitro-aortic homograft valve replacement was performed in 29 cases. Results: Mean follow-up was 9.8 ± 6.3 years (up to 19.2 yrs). There were 7 early (2
文摘Aim: To 1) compare post-wash and post-thaw parameters of sperm processed with PureSperm density gradient technique and swim-up method; and 2) test the efficacy of two commonly available density gradient media PureSperm and ISolate. Methods: This prospective study used semen specimens from 22 patients. Specimens from nine patients were processed by both PureSperm density gradient and swim-up method. These specimens were then cryopreserved. Thirteen specimens were processed by both PureSperm (40 % and 80 %) and Isolate (50 % and 90 %) double density gradient techniques. The two fractions processed by both PureSperm and swim-up were analyzed for post-wash sperm characteristics. Post-thaw analysis was done after 24 hours. Sperm fractions obtained after processing with PureSperm and ISolate were compared for post-wash sperm characteristics and ROS levels. Results: Specimens prepared with PureSperm had significantly higher median total motile sperm counts (TMSC) (32.2 x 10~6 vs. 17.6 x 10~6), recovery rates (69.2 % vs. 50.0 %), and longevity at 4 hours (83.0 % vs. 55.0 %) compared to specimen prepared by swim-up. Post-thaw specimens also had a higher recovery and longevity at 4 hours with PureSperm as compared to the swim-up. Semen specimens processed by PureSperm had significantly higher total sperm count, TMSC, and percentage recovery rates (30.0 % vs. 19.7 %) than ISolate. Conclusion: Semen quality is better preserved in fresh and cryopreserved semen prepared with PureSperm density gradient compared to swim-up. A significant enrichment of sperm is observed with PureSperm compared to ISolate. Higher recovery rates of mature motile sperm obtained after PureSperm sperm preparation may be beneficial for successful ART. i
基金Project supported by the Science Foundation (No. 2003C23015) and the Natural Science Foundation (No. Y204139) of Zhejiang Province, China
文摘This study is aimed at investigating the potentials of ex vivo expansion and pluri-differentiation of cryopreservation of adult human bone marrow mesenchymal stem cells （hMSCs） into chondrocytes, adipocytes and neurocytes. Cryopreserved hMSCs were resuscitated and cultured for 15 passages, and then induced into chondrocytes, adipocytes and neurocytes with corresponding induction medium. The induced cells were observed for morphological properties and detected for expressions of type II collagen, triglyceride or neuron-specific enolase and nestin. The result showed that the resuscitated cells could differentiate into chondrocytes after exposure to transforming growth factor 61 （TGF-~0, insulin-like growth factor I （IGF-I） and vitamin C （Vc）, and uniformly changed morphologically from a spindle-like fibroblastic appearance to a polygonal shape in three weeks. The induced cells were heterochromatic to safranin O and expressed cartilage matrix-procollagenal （If） mRNA. The resuscitated cells cultured in induction medium consisting of dexamethasone, 3-isobutyl-l-methylxanthine, indomethacin and IGF-I showed adipogenesis, and lipid vacuoles accumulation was detectable after 21 d. The resuscitated hMSCs were also induced into neurocytes and expressed nestin and neuron specific endolase （NSE） that were special surface markers associated with neural cells at different stage. This study suggested that the resuscitated hMSCs should be still a population ofpluripotential cells and that it could be used for establishing an abundant bMSC reservoir for further experiment and treatment of various clinical discases.
文摘Cryopreservation is associated with the production of reactive oxygen species, which leads to lipid peroxidation of the sperm membrane and consequently a reduction in sperm motility and decreased fertility potential. The aim of this study was to determine the optimal concentration of L-cysteine needed for cryopreservation of boar semen. Twelve boars provided semen of proven motility and morphology for this study. The semen was divided into four portions in which the lactose-egg yolk （LEY） extender used to resuspend the centrifuged sperm pellet was supplemented with various concentrations of L-cysteine to reach 0 mmol L^-1 （group Ⅰ, control）, 5 mmol L^-1 （group Ⅱ）, 10 mmol L^-1 （group Ⅲ） and 15 mmol L^-1 （group Ⅳ）. Semen suspensions were loaded in straws （0.5 mL） and placed in a controlled-rate freezer. After cryopreservation, frozen semen samples were thawed and investigated for progressive motility, viability using SYBR-14/EthD-1 staining and acrosome integrity using FITC-PNA/EthD-1 staining. There was a significantly higher （P 〈 0.01） percentage of progressive motility, viability and acrosomal integrity in two L-cysteine-supplemented groups （group Ⅱ and group Ⅲ） compared with the control. There was a biphasic effect of L-cysteine, with the highest percentage of progressive motility, viability and acrosomal integrity in group Ⅲ. In conclusion, 5 or 10 mmol L^-1 was the optimum concentration of L-cysteine to be added to the LEY extender for improving the quality of frozen-thawed boar semen.
基金This research was sponsored by the National Key Research and Development Projects(No.2018YFC1003600,2016YFC1000302,2017YFC1002001 and SQ2018YFC100243)the Clinical Medicine PlusX Young Scholars Project,Peking University(No.2102018237)the Beijing Municipal Natural Science Foundation(No.7182177).
文摘We performed this study to evaluate the clinical outcomes of microdissection testicular sperm extraction-intracytoplasmic sperm injection(micro-TESE-ICSI)treatment that used fresh or cryopreserved sperm in patients with nonobstructive azoospermia(NOA).A total of 338 NOA patients with 344 consecutive cycles received treatment in the reproductive medicine center of Peking University Third Hospital in Beijing,China,from January 2014 to December 2017.Fresh oocytes and fresh sperm were used in 222 patients with 234 cycles(Group A).Fresh oocytes and cryopreserved sperm were used in 116 patients with 110 cycles(Group B).We compared patient characteristics,embryonic development,and pregnancy outcomes between Groups A and B.There was no statistical difference in the patient characteristics,and no differences were observed with fertilization or quality embryo rates between Groups A and B.The rates of clinical pregnancy and live birth were both higher for Group A than those for Group B(both P<0.05).In conclusion,fresh testicular sperm appears to produce better ICSI outcomes than cryopreserved testicular sperm in patients with NOA.
基金supported by Young Investigator Research Promotion Grant, Chulalongkorn Uni- versity
文摘The aim of this article was to examine the research articles regarding biological and mechanical properties of cryopreserved teeth for potential use in tooth transplantation. A systematic review of literatures was performed by Pubmed searching with assigned key words from January 1, 1990 to June 8, 2009. All articles were examined for inclusion criteria. Secondary search was conducted by hand-search through references of included articles from primary search. A total of 24 articles were obtained from both primary and secondary search and used as fundamental articles in this review. Periodontal ligament tissues of cryopreserved teeth were able to maintain their biological properties resulted in a satisfactory healing of periodontium. Dental pulp tissues,however, may be compromised by limitation of permeability of cryopreservative agent into pulp cavity. Therefore, an endodontic treatment of transplanted cryopreserved teeth was recommended. Cryopreserved teeth had comparable mechanical properties to those of normal teeth. Importantly, the success of cryopreserved tooth transplantation treatment in orthodontic patients was reported. The cryopreserved teeth for tooth banking have a potential clinical application for treatment of missing teeth. Case selection, however, is critical for treatment success. More studies and data regarding masticatory function and periodontal healing of transplanted cryopreserved teeth are needed.
基金Project supported by the Major Program for Science and Technology Research of Zhejiang Province(No.2011C13029-2)the Medical Scientific Research Foundation of Zhejiang Province(Nos2012ZDA026 and 2013ZDA012),China
文摘Objective： To evaluate the clinical efficacy of peripheral deep anterior lamellar keratoplasty （DALK） using a cryopreserved donor cornea for Terrien＇s marginal degeneration （TMD）. Methods： Thirty-one eyes of 27 patients with TMD underwent peripheral DALK using cryopreserved donor corneas, According to the distance between the inner edge of the lesion and the limbus, a ring-shaped or D-shaped DALK was performed. All grafts were stored at -20 ℃. Cryopreserved comeoscleral rims were prepared for ring-shaped grafts and cryopreserved whole eyeballs were prepared for D-shaped grafts. The general conditions, intraoperative performance, postoperative corneal reconstruc- tion, astigmatism, best corrected visual acuity （BCVA）, and various complications were analyzed. Results： Ring-shaped DALK was performed in 28 eyes and D-shaped DALK was performed in 3 eyes. Postoperative follow-up time was （28.4±24.8） months. There was evidence of inflammation before surgery in 12 eyes （38.7%） and intraoper- ative perforation occurred in 13 eyes （41.9%）. The corneal structures of all eyes were reconstructed. Postoperative astigmatism and BCVA showed improvement （both P=0.00） except for cases that underwent D-shaped DALK. Ten eyes （32.2%） developed transient ocular hypertension and one eye （3.2%） developed secondary glaucoma. No pri- mary disease recurrence or corneal allograft rejection was observed. Conclusions： Peripheral DALK for TMD using cryopreserved donor tissue is an effective technique that eliminates rejection and extends the use of donor eyes. Inflammatory history or intraoperative perforation has no adverse effect on graft recovery. However, D-shaped DALK did not achieve good visual outcomes.
基金supported by the National Natural Science Foundation of China,No.81201546the Doctoral Start-up Program of Natural Science Foundation of Guangdong Province of China,No.2017A030310302+1 种基金the Medical Scientific Research Foundation of Guangdong Province of China,No.A2016018grants from the Science and Technology Project of Guangdong Province of China,No.2016A010103012,2013B010404019
文摘The experimental design evaluated histological,mechanical,and biological properties of allogeneic decellularized nerves after cryopreservation in a multi-angle,multi-directional manner to provide evidence for long-term preservation.Acellular nerve allografts from human and rats were cryopreserved in a cryoprotectant（10% fetal bovine serum,10% dimethyl sulfoxide,and 5% sucrose in RPMI1640 medium） at-80°C for 1 year,followed by thawing at 40°C or 37°C for 8 minutes.The breaking force of acellular nerve allografts was measured using a tensile test.Cell survival was determined using L-929 cell suspensions.Acellular nerve allografts were transplanted into a rat model with loss of a 15-mm segment of the left sciatic nerve.Immunohistochemistry staining was used to measure neurofilament 200 expression.Hematoxylin-eosin staining was utilized to detect relative muscle area in gastrocnemius muscle.Electron microscopy was applied to observe changes in allograft ultrastructure.There was no obvious change in morphological appearance or ultrastructure,breaking force,or cytotoxicity of human acellular nerve allografts after cryopreservation at-80°C.Moreover,there was no remarkable change in neurofilament 200 expression,myelin sheath thickness,or muscle atrophy when fresh or cryopreserved rat acellular nerve allografts were applied to repair nerve injury in rats.These results suggest that cryopreservation can greatly extend the storage duration of acellular nerve tissue allografts without concomitant alteration of the physiochemical and biological properties of the engineered tissue to be used for transplantation.
文摘The relative merit of four different methods (method of fine tube, semen pellets, ampule, and test tube)of cryopreserved spermatozoa from 43 male donors with normal fertile ability was evaluated.The mobility and viability in post-thawed 172 samples were examined. The method of fine tube was recommedned to be the method of choice. A discussion on these four methods was given.
文摘Objective： To study the method of cryopreserving rat hepatocytes and double collagen gel culture measurement after its cryopreservation. Methods： Rat hepatocytes, isolated by two-step perfusion with collagenase using an extra corporeal perfusion apparatus, were cryopreserved in double collagen gel with culture medium added by epidermal growth factor（EGF）. The expression of cell function and cellular morphology were examined during culture. Results： The hepatocytes cryopreserved in double collagen gel concluding EGF showed good morphology and biological characteristics. After thawing, the MTT metabolism and protein synthesis of hepatocytes in sandwich ± EGF groups were better than those in control group. And the morphology and function of hepatocytes in sandwich group was better than that in EGF group（P 〈 0.05）. Conclusion： Double collagen gel culture can keep hepatocyte＇s activities. Thawed hepatocytes can be cultivated with collagenous matrix, which provides an environment that more closely resembles that in vivo and maintain the expression of certain liver-specific function of hepatocytes.
基金Supported by National Natural Science Foundation of China(No.81300736No.81370993)
文摘This study aimed to evaluate the outcomes and described the recovery process of cryopreserved limbal lamellar keratoplasty（CLLK） for peripheral corneal and limbal diseases. Thirteen eyes of 12 patients with a mean age of 41±23.9 y were included. The average follow-up was 12.1±5.6 mo. Stable ocular surface was achieved in all eyes at last follow-up. Epithelialization originated from both recipient and graft in 9 eyes. We conclude that CLLK compensates for the shortage of donor corneas and cryopreserved limbal grafts provide epithelialization sources in ocular surface reconstruction.
文摘Objective:To test the effects of extenders and packaging methods on morphological and functional characteristics of frozen thawed Ossimi ram semen.Methods:Ram semen was pooled,diluted in 3 different extenders:Tris-egg yolk(TEY),Tris-soybean lecithin(TSBL),and Tris-butylatedhydroxytoluene(TBHT),equilibrated at 5℃for 4 h,and packaged in straws or pellets for freezing.Semen was evaluated for sperm progressive motility,viability,abnormality,and membrane integrity after dilution,equilibration and thawing.The percentages of viable,early apoptotic,apoptotic,and necrotic spermatozoa as well as comet assay parameters were determined in post-thawed semen.Total antioxidants capacity,malondialdehyde and lactic dehydrogenase were assayed in thawed seminal plasma.Results:After equilibration,only sperm membrane integrity was significantly higher(P<0.05)in TEY and TSBL than in TBHT.After thawing,TEY or TSBL in straws significantly improved sperm progressive motility and vitality(P<0.05).In thawed seminal plasma,TBHT or TSBL in straws and TBHT in pellets significantly reduced malondialdehyde,and TBHT in pellets significantly increased lactic dehydrogenase(P<0.05).TEY in straws increased viable sperm,while significantly decreased early apoptotic and apoptotic sperm(P<0.05).DNA damage was significantly decreased(P<0.05)in straws with TEY and TSBL,and tail moment decreased in straws with all extenders(P<0.05).Conclusions:Despite the disadvantages of dilution of cryopreserved semen with egg yolk,ram semen cryopreserved with TEY gives the best physical,morphological and functional characteristics in straws compared with pellets,followed by semen diluted with TSBL.However,semen diluted with TBHT or TSBL,regardless of packaging method,showed the highest impact on antioxidant status of cyopreserved ram semen.
