Mesenchymal stem cells （MSCs） show the great promise for the treatment of a variety of diseases because of their self-renewal and multipotential abilities. MSCs are generally cultured on two-dimensional （2D） subst...Mesenchymal stem cells （MSCs） show the great promise for the treatment of a variety of diseases because of their self-renewal and multipotential abilities. MSCs are generally cultured on two-dimensional （2D） substrate in vitro. There are indications that they may simultaneously lose their sternness and multipotentiality as the result of prolonged 2D culture. In this study, we used three-dimensional （3D） collagen scaffolds as rat MSCs cartier and compared the properties of MSCs on 3D collagen scaffolds with monolayer cultured MSCs. The results demonstrated that collagen scaffolds were suitable for rat MSCs adherence and proliferation. More importantly, compared to MSCs under 2D culture, 3D MSCs significantly maintained higher expression levels of stemness genes （Oct4, Sox2, Rex-1 and Nanog）, yielded high frequencies of colony-forming units-fibroblastic （CFU-F） and showed enhanced osteogenic and adipogenic differentiation efficiency upon induction. Thus, 3D collagen scaffolds may be beneficial for expanding rat MSCs while maintaining the stem cell properties in vitro.展开更多
This study aimed to investigate the neural differentiation of human umbilical cord mesenchymal stem cells （hUCMSCs） under the induction of injured neural cells. After in vitro isolation and culture, passage 5 hUCMSC...This study aimed to investigate the neural differentiation of human umbilical cord mesenchymal stem cells （hUCMSCs） under the induction of injured neural cells. After in vitro isolation and culture, passage 5 hUCMSCs were used for experimentation, hUCMSCs were co-cultured with normal or AI31.4o-injured PC12 cells, PC12 cell supernatant or PC12 cell lysate in a Transwell co-culture system. Western blot analysis and flow cytometry results showed that choline acetyltransferase and microtubule-associated protein 2, a specific marker for neural cells, were expressed in hUCMSCs under various culture conditions, and highest expression was observed in the hUCMSCs co-cultured with injured PC12 cells. Choline acetyltransferase and microtubule-associated protein 2 were not expressed in hUCMSCs cultured alone （no treatment）. Cell Counting Kit-8 assay results showed that hUCMSCs under co-culture conditions promoted the proliferation of injured PC12 cells. These findings suggest that the microenvironment during neural tissue injury can effectively induce neural cell differentiation of hUCMSCs. These differentiated hUCMSCs likely accelerate the repair of injured neural ceils.展开更多
Poor quality embryos discarded from in vitro fertilization （IVF） laboratories are good sources for deriving human embryonic stem cell （hESC） lines. In this study, 166 poor quality embryos donated from IVF centers ...Poor quality embryos discarded from in vitro fertilization （IVF） laboratories are good sources for deriving human embryonic stem cell （hESC） lines. In this study, 166 poor quality embryos donated from IVF centers on day 3 were cultured in a blastocyst medium for 2 days, and 32 early blastocysts were further cultured in a blastocyst optimum culture medium for additional 2 days so that the inner cell masses （ICMs） could be identified and isolated easily. The ICMs of 17 blastocysts were isolated by a mechanical method, while those of the other 15 blastocysts were isolated by immunosurgery. All isolated ICMs were inoculated onto a feeder layer for subcultivation. The rates of ICM attachment, primary ICM colony formation and the efficiency of hESC derivation were similar between the ICMs isolated by the two methods （P〉0.05）. As a result, four new hESC lines were established. Three cell lines had normal karyotypes and one had an unbalanced Robertsonian translocation. All cell lines showed normal hESC characteristics and had the differentiation ability. In conclusion, we established a stable and effective method for hESC isolation and culture, and it was confirmed that the mechanical isolation was an effective method to isolate ICMs from poor embryos. These results further indicate that hESC lines can be derived from poor quality embryos discarded by IVF laboratories.展开更多
Objective： Recent evidence suggests that Oct-4 is highly expressed in several cancers, and its expression contributes to tumor growth. In this study, we investigated the level of Oct-4 expression in rectal adenocarci...Objective： Recent evidence suggests that Oct-4 is highly expressed in several cancers, and its expression contributes to tumor growth. In this study, we investigated the level of Oct-4 expression in rectal adenocarcinoma, and evaluated the prognostic significance of Oct-4 expression in these cases. Methods： The immunohistochemical expression of Oct-4 was evaluated in 52 formalin-fixed paraffin-embedded postoperative rectal adenocarcinoma tissue samples. The impact of the immunoreactivity of Oct-4 in regard to clinical outcome was determined by Kaplan-Meier and log-rank. Results： The expression level of Oct-4 ranged from 0 to 18.5%. There was no significant association between Oct-4 expression and gender （P=0.772）, age （P=0.123）, clinical stage （P=0.391）, and histological grade （P=0.056）. The 3-year local recurrence-free rates with negative and positive expression of Oct-4 were 83.5% and 75.0%, respectively （P=0.583）. The 3-year metastasis-free rates with negative and positive expression of Oct-4 were 88.6% and 61.9%, respectively （P=0.035）. The 3-year overall survival rates with negative and positive expression of Oct-4 were 77.9% and 49.0%, respectively （P=0.037）. Conclusion： The results suggest that embryonic stem cell marker Oct-4 expression may have prognostic significance in patients with rectal adenocarcinoma. However, to confirm this more and larger studies are required.展开更多
Our preliminary studies confirmed that an active principle region of Buyang Huanwu decoction, comprising alkaloid, polysaccharide, aglycon, glucoside and volatile oil, can induce bone marrow mesenchymal stem cell diff...Our preliminary studies confirmed that an active principle region of Buyang Huanwu decoction, comprising alkaloid, polysaccharide, aglycon, glucoside and volatile oil, can induce bone marrow mesenchymal stem cell differentiation into neurons. Mitogen-activated protein kinase signaling was identified as one of the key pathways underlying this differentiation process. The present study shows phosphorylated extracellular signal-regulated protein kinase and phosphorylated p38 protein expression was increased after differentiation. Cellular signaling pathway blocking agents, PD98059 and SB203580, inhibited extracellular signal-regulated protein kinase and p38 in mitogen-activated protein kinase signaling pathways respectively, mRNA and protein expression of the neuronal marker, neuron specific enolase, and neural stem cell marker, nestin, were decreased in bone marrow mesenchymal stem cells after treatment with the active principle region of Buyang Huanwu decoction. Experimental findings indicate that, extracellular signal-regulated protein kinase and p38 in mitogen-activated protein kinase signaling pathways participate in bone marrow mesenchymal stem cell differentiation into neuron-like cells, induced by the active principle region of Buyang Huanwu decoction.展开更多
A novel fimctional gene P12 was isolated from neural stem cells cultured in differentiation medium. The fully length cDNA of P12 gene was cloned and sequenced. Result showed that it contains an open reading frame enco...A novel fimctional gene P12 was isolated from neural stem cells cultured in differentiation medium. The fully length cDNA of P12 gene was cloned and sequenced. Result showed that it contains an open reading frame encoding a protein of 291 amino acids. Further, this gene was transferred into neural stem cell. Functional analysis suggests that the expression of p12 protein is closely correlated with differentiation of neural dendrite configuration. In addition, to obtain encoding protein, P12 sequence was also expressed in Pichia pastoris yeast.展开更多
Objective： Colon cancer is one of the most common human malignancies. Cancer stem cells （CSCs）, despite being only a small subset of cancer cells, have the capability to self-renew and sustain the tumor. They also ...Objective： Colon cancer is one of the most common human malignancies. Cancer stem cells （CSCs）, despite being only a small subset of cancer cells, have the capability to self-renew and sustain the tumor. They also have the ability to proliferate. Multiple CSCs-associated markers have been identified in colon cancer including CD133, ALDH1 and β-catenin. The aim of the work was to study the prognostic value of CSCs markers （CD133, ALDH 1 and β-catenin）, as well as their rela- tionship to clinicopathological features of colon cancer. Methods： CD133, ALDH1 and β-catenin proteins expression was as- sessed immunohistochemically in a series of colon cancers and their prognostic significance was evaluated. Results： CD133 expression showed significant relationship to tumor stage and lymph node metastasis （P-value 0.004 ＆ 〈 0.001 respectively）, and near significant relationship to liver metastasis （P-value 0.092）. ALDH1 was significantly associated with tumor grade, stage and nodal metastasis （P-value 0.021,0.001 and 0.026 respectively）, but its relationship to liver metastasis was near sig- nificant （P-value 0.068）. Nuclear β-catenin was significantly related to tumor grade, stage, nodal and liver metastasis （P-value 0.001, 〈 0.001, 〈 0.001 and 0.008 respectively）. Overall survival （OS） was associated inversely with CD133, ALDH1 positivity, and directly with nuclear 13-catenin posiUvity （P-value 〈 0.001,0.0001 and 〈 0.001 respectively）. Also recurrence free survival （RFS） was associated inversely with CD133, ALDH1 and directly with nuclearβ-catenin positivity （P-value 0.0001,0.001 and 〈 0.001 respectively）. Conclusion： CD133, ALDH1 and β-catenin expressions of tumor cells have significant impact upon malignant progression of colon cancer and thus patient survival and tumor recurrence. Hence they can be used to predict outcome of colon cancer patients.展开更多
Ginsenoside Rgl is the major pharmacologically active component of ginseng, and is reported to have various therapeutic actions. To determine whether it induces the differentiation of neural stem cells, and whether ne...Ginsenoside Rgl is the major pharmacologically active component of ginseng, and is reported to have various therapeutic actions. To determine whether it induces the differentiation of neural stem cells, and whether neural stem cell transplantation after induction has therapeutic effects on hypoxic-ischemic encephalopathy, we cultured neural stem cells in 10-80 ~tM ginsenoside Rgl. Immunohistochemistry revealed that of the concentrations tested, 20 mM ginsenoside Rgl had the greatest differentiation-inducing effect and was the concentration used for subsequent exper- iments. Whole-cell patch clamp showed that neural stem cells induced by 20 jaM ginsenoside Rgl were more mature than non-induced cells. We then established neonatal rat models of hypox- ic-ischemic encephalopathy using the suture method, and ginsenoside Rgl-induced neural stem cells were transplanted via intracerebroventricular injection. These tests confirmed that neural stem cells induced by ginsenoside had fewer pathological lesions and had a significantly better behavioral capacity than model rats that received saline. Transplanted neural stem cells expressed neuron-specific enolase, and were mainly distributed in the hippocampus and cerebral cortex. The present data suggest that ginsenoside Rgl-induced neural stem cells can promote the partial recovery of complicated brain functions in models of hypoxic-ischemic encephalopathy.展开更多
基金supported by the grants from the Ministry of Science and Technology of China(Nos.2011CB965001 and 2011CB710905)the Knowledge Innovation Program of the Chinese Academy of Sciences(Nos.KSCX2-YW-R-232, KJCX2-YW-L08 and KYQY-QN-015)
文摘Mesenchymal stem cells （MSCs） show the great promise for the treatment of a variety of diseases because of their self-renewal and multipotential abilities. MSCs are generally cultured on two-dimensional （2D） substrate in vitro. There are indications that they may simultaneously lose their sternness and multipotentiality as the result of prolonged 2D culture. In this study, we used three-dimensional （3D） collagen scaffolds as rat MSCs cartier and compared the properties of MSCs on 3D collagen scaffolds with monolayer cultured MSCs. The results demonstrated that collagen scaffolds were suitable for rat MSCs adherence and proliferation. More importantly, compared to MSCs under 2D culture, 3D MSCs significantly maintained higher expression levels of stemness genes （Oct4, Sox2, Rex-1 and Nanog）, yielded high frequencies of colony-forming units-fibroblastic （CFU-F） and showed enhanced osteogenic and adipogenic differentiation efficiency upon induction. Thus, 3D collagen scaffolds may be beneficial for expanding rat MSCs while maintaining the stem cell properties in vitro.
文摘This study aimed to investigate the neural differentiation of human umbilical cord mesenchymal stem cells （hUCMSCs） under the induction of injured neural cells. After in vitro isolation and culture, passage 5 hUCMSCs were used for experimentation, hUCMSCs were co-cultured with normal or AI31.4o-injured PC12 cells, PC12 cell supernatant or PC12 cell lysate in a Transwell co-culture system. Western blot analysis and flow cytometry results showed that choline acetyltransferase and microtubule-associated protein 2, a specific marker for neural cells, were expressed in hUCMSCs under various culture conditions, and highest expression was observed in the hUCMSCs co-cultured with injured PC12 cells. Choline acetyltransferase and microtubule-associated protein 2 were not expressed in hUCMSCs cultured alone （no treatment）. Cell Counting Kit-8 assay results showed that hUCMSCs under co-culture conditions promoted the proliferation of injured PC12 cells. These findings suggest that the microenvironment during neural tissue injury can effectively induce neural cell differentiation of hUCMSCs. These differentiated hUCMSCs likely accelerate the repair of injured neural ceils.
基金the Guangdong Province Health Department (No. B30202)Guangzhou City Science and Technology Administration (No. 2006Z1-E0021)+1 种基金partly supported by the Guangdong Province Science and Technology Administration (No. 2007A032100003)Guangdong Provincial Medical Research Fund (No. A2008287)
文摘Poor quality embryos discarded from in vitro fertilization （IVF） laboratories are good sources for deriving human embryonic stem cell （hESC） lines. In this study, 166 poor quality embryos donated from IVF centers on day 3 were cultured in a blastocyst medium for 2 days, and 32 early blastocysts were further cultured in a blastocyst optimum culture medium for additional 2 days so that the inner cell masses （ICMs） could be identified and isolated easily. The ICMs of 17 blastocysts were isolated by a mechanical method, while those of the other 15 blastocysts were isolated by immunosurgery. All isolated ICMs were inoculated onto a feeder layer for subcultivation. The rates of ICM attachment, primary ICM colony formation and the efficiency of hESC derivation were similar between the ICMs isolated by the two methods （P〉0.05）. As a result, four new hESC lines were established. Three cell lines had normal karyotypes and one had an unbalanced Robertsonian translocation. All cell lines showed normal hESC characteristics and had the differentiation ability. In conclusion, we established a stable and effective method for hESC isolation and culture, and it was confirmed that the mechanical isolation was an effective method to isolate ICMs from poor embryos. These results further indicate that hESC lines can be derived from poor quality embryos discarded by IVF laboratories.
基金supported by the grant from Jiangsu Natural Science Program(No.BK2009126).
文摘Objective： Recent evidence suggests that Oct-4 is highly expressed in several cancers, and its expression contributes to tumor growth. In this study, we investigated the level of Oct-4 expression in rectal adenocarcinoma, and evaluated the prognostic significance of Oct-4 expression in these cases. Methods： The immunohistochemical expression of Oct-4 was evaluated in 52 formalin-fixed paraffin-embedded postoperative rectal adenocarcinoma tissue samples. The impact of the immunoreactivity of Oct-4 in regard to clinical outcome was determined by Kaplan-Meier and log-rank. Results： The expression level of Oct-4 ranged from 0 to 18.5%. There was no significant association between Oct-4 expression and gender （P=0.772）, age （P=0.123）, clinical stage （P=0.391）, and histological grade （P=0.056）. The 3-year local recurrence-free rates with negative and positive expression of Oct-4 were 83.5% and 75.0%, respectively （P=0.583）. The 3-year metastasis-free rates with negative and positive expression of Oct-4 were 88.6% and 61.9%, respectively （P=0.035）. The 3-year overall survival rates with negative and positive expression of Oct-4 were 77.9% and 49.0%, respectively （P=0.037）. Conclusion： The results suggest that embryonic stem cell marker Oct-4 expression may have prognostic significance in patients with rectal adenocarcinoma. However, to confirm this more and larger studies are required.
基金sponsored by the National Natural Science Foundation of China,No.81102595the Natural Science Foundation of Guangxi,No.2012GXNSFAA053113
文摘Our preliminary studies confirmed that an active principle region of Buyang Huanwu decoction, comprising alkaloid, polysaccharide, aglycon, glucoside and volatile oil, can induce bone marrow mesenchymal stem cell differentiation into neurons. Mitogen-activated protein kinase signaling was identified as one of the key pathways underlying this differentiation process. The present study shows phosphorylated extracellular signal-regulated protein kinase and phosphorylated p38 protein expression was increased after differentiation. Cellular signaling pathway blocking agents, PD98059 and SB203580, inhibited extracellular signal-regulated protein kinase and p38 in mitogen-activated protein kinase signaling pathways respectively, mRNA and protein expression of the neuronal marker, neuron specific enolase, and neural stem cell marker, nestin, were decreased in bone marrow mesenchymal stem cells after treatment with the active principle region of Buyang Huanwu decoction. Experimental findings indicate that, extracellular signal-regulated protein kinase and p38 in mitogen-activated protein kinase signaling pathways participate in bone marrow mesenchymal stem cell differentiation into neuron-like cells, induced by the active principle region of Buyang Huanwu decoction.
基金Project supported by National Basic Research Program of China(Grant No . 973 -2006CB500702) , National Natural Science Foundation of China (Grant No .30570590) ,Science Foundation of Shanghai Municipal Commission of Science and Technology(Grant Nos . 03JC1403 , 03JC14061) , and Scientific Research Foundation of the Returned Overseas Chinese Scholars , State Education Ministry (Grant No .JWSL2003406)
文摘A novel fimctional gene P12 was isolated from neural stem cells cultured in differentiation medium. The fully length cDNA of P12 gene was cloned and sequenced. Result showed that it contains an open reading frame encoding a protein of 291 amino acids. Further, this gene was transferred into neural stem cell. Functional analysis suggests that the expression of p12 protein is closely correlated with differentiation of neural dendrite configuration. In addition, to obtain encoding protein, P12 sequence was also expressed in Pichia pastoris yeast.
文摘Objective： Colon cancer is one of the most common human malignancies. Cancer stem cells （CSCs）, despite being only a small subset of cancer cells, have the capability to self-renew and sustain the tumor. They also have the ability to proliferate. Multiple CSCs-associated markers have been identified in colon cancer including CD133, ALDH1 and β-catenin. The aim of the work was to study the prognostic value of CSCs markers （CD133, ALDH 1 and β-catenin）, as well as their rela- tionship to clinicopathological features of colon cancer. Methods： CD133, ALDH1 and β-catenin proteins expression was as- sessed immunohistochemically in a series of colon cancers and their prognostic significance was evaluated. Results： CD133 expression showed significant relationship to tumor stage and lymph node metastasis （P-value 0.004 ＆ 〈 0.001 respectively）, and near significant relationship to liver metastasis （P-value 0.092）. ALDH1 was significantly associated with tumor grade, stage and nodal metastasis （P-value 0.021,0.001 and 0.026 respectively）, but its relationship to liver metastasis was near sig- nificant （P-value 0.068）. Nuclear β-catenin was significantly related to tumor grade, stage, nodal and liver metastasis （P-value 0.001, 〈 0.001, 〈 0.001 and 0.008 respectively）. Overall survival （OS） was associated inversely with CD133, ALDH1 positivity, and directly with nuclear 13-catenin posiUvity （P-value 〈 0.001,0.0001 and 〈 0.001 respectively）. Also recurrence free survival （RFS） was associated inversely with CD133, ALDH1 and directly with nuclearβ-catenin positivity （P-value 0.0001,0.001 and 〈 0.001 respectively）. Conclusion： CD133, ALDH1 and β-catenin expressions of tumor cells have significant impact upon malignant progression of colon cancer and thus patient survival and tumor recurrence. Hence they can be used to predict outcome of colon cancer patients.
基金supported by the Natural Science Foundation of Chongqing in China,No.CSTC2011jj A0013
文摘Ginsenoside Rgl is the major pharmacologically active component of ginseng, and is reported to have various therapeutic actions. To determine whether it induces the differentiation of neural stem cells, and whether neural stem cell transplantation after induction has therapeutic effects on hypoxic-ischemic encephalopathy, we cultured neural stem cells in 10-80 ~tM ginsenoside Rgl. Immunohistochemistry revealed that of the concentrations tested, 20 mM ginsenoside Rgl had the greatest differentiation-inducing effect and was the concentration used for subsequent exper- iments. Whole-cell patch clamp showed that neural stem cells induced by 20 jaM ginsenoside Rgl were more mature than non-induced cells. We then established neonatal rat models of hypox- ic-ischemic encephalopathy using the suture method, and ginsenoside Rgl-induced neural stem cells were transplanted via intracerebroventricular injection. These tests confirmed that neural stem cells induced by ginsenoside had fewer pathological lesions and had a significantly better behavioral capacity than model rats that received saline. Transplanted neural stem cells expressed neuron-specific enolase, and were mainly distributed in the hippocampus and cerebral cortex. The present data suggest that ginsenoside Rgl-induced neural stem cells can promote the partial recovery of complicated brain functions in models of hypoxic-ischemic encephalopathy.
