Objective To study large-scale expansion of SD （Sprague-Dawley） rat＇s osteoblasts in suspension culture in a rotating wall vessel bioreactor （RWVB）. Methods The bioreactor rotation speeds were adjusted in the ran...Objective To study large-scale expansion of SD （Sprague-Dawley） rat＇s osteoblasts in suspension culture in a rotating wall vessel bioreactor （RWVB）. Methods The bioreactor rotation speeds were adjusted in the range of 0 to 20 rpm, which could provide low shear on the rnicrocarriers around 1 dyn/cm^2. The cells were isolated via sequential digestions of neonatal （less than 3 days old） SD rat calvaria. After the primary culture and several passages, the cells were seeded onto the microcarriers and cultivated in T-flask, spinner flask and RWVB respectively. During the culture period, the cells were counted and observed under the inverted microscope for morphology every 12 h. After 7 days, the cells were evaluated with scanning electron microscope （SEM） for histological examination of the aggregates. Also, the hematoxylin-eosin （HE） staining and alkaline phosphatase （ALP） staining were performed. Moreover, von-Kossa staining and Alizarin Red S staining were carded out for mineralized nodule formation. Results The results showed that in RWVB, the cells could be expanded by more than ten times and they presented better morphology and vitality and stronger ability to form bones. Conclusions The developed RWVB can provide the culture environment with a relatively low shear force and necessary three-dimensional （3D） interactions among cells and is suitable for osteopath expansion in vitro.展开更多
Objective To analyze the forces of rotational wall vessel （RWV） bioreactor on small tissue pieces or microcarrier particles and to determine the tracks of microcarrier particles in RWV bioreactor. Methods The motion...Objective To analyze the forces of rotational wall vessel （RWV） bioreactor on small tissue pieces or microcarrier particles and to determine the tracks of microcarrier particles in RWV bioreactor. Methods The motion of the microcarrier in the rotating wall vessel （RWV） bioreactor with both the inner and outer cylinders rotating was modeled by numerical simulation. Results The continuous trajectory of microcarrier particles, including the possible collision with the wall was obtained. An expression between the minimum rotational speed difference of the inner and outer cylinders and the microcarrier particle or aggregate radius could avoid collisions with either wall. The range of microcarrier radius or tissue size, which could be safely cultured in the RWV bioreactor, in terms of shear stress level, was determined. Conclusion The model works well in describing the trajectory of a heavier microcarrier particle in rotating wall vessel.展开更多
1 IntroductionRecent research shows that neural stem cells mayplay an important role in the nerve injury reparation andnerve disease treatment.The shortage of the source andthe number of NSCs,however,is the main chall...1 IntroductionRecent research shows that neural stem cells mayplay an important role in the nerve injury reparation andnerve disease treatment.The shortage of the source andthe number of NSCs,however,is the main challenge forits clinic application.In thi s situation,expansion of NSCsin large scale and culture in three dimensional enviorrment are very worth of exploration.Notablely,the shearstress existed in bioreactors can cause serious cell injuryespecially for the shear sensitive cells like NSCs.展开更多
1 IntroductionAnogeneic bone marrow transplantation is used forthe treatment of a variety of malignant and mnmalignanthematological disorders.Human umbilical cord blood isanother source of hematopoietic stem cells.It ...1 IntroductionAnogeneic bone marrow transplantation is used forthe treatment of a variety of malignant and mnmalignanthematological disorders.Human umbilical cord blood isanother source of hematopoietic stem cells.It is acceptedthat there are definite advantages in the use of cord blood(CB)as a source of haematqpoietic progenitor cells(HPC)in selected patients with malignant and mr malig-nant haematological conditions.展开更多
Introduction1Vitrification is an effective method for cryopreservation of cell s[1.2].Hbwever,cells are usually damageddue to the osmotic injuy caused by the higher concentra-tions of cryoprotective agents(CPA)during ...Introduction1Vitrification is an effective method for cryopreservation of cell s[1.2].Hbwever,cells are usually damageddue to the osmotic injuy caused by the higher concentra-tions of cryoprotective agents(CPA)during CPA remov-ing.The ice recrystallization in thawing solution can alsohurt cells serio usly.Antifreeze glycoprotein(AFGPs)isextremely efficient at inhibiting ice recrystallization[3].展开更多
1 IntroductionClinical trials have demonstrated that ex vivo ex-panded hematopoietic stem cells(HSCs)and progenitorsoffer great pomise in reconstituting in vivo hematopoiesisin patients who lave undergpne intensive ch...1 IntroductionClinical trials have demonstrated that ex vivo ex-panded hematopoietic stem cells(HSCs)and progenitorsoffer great pomise in reconstituting in vivo hematopoiesisin patients who lave undergpne intensive chemmtherapy.It is therefore necessary to develop a clinical scale culturesystem to provide the expanded HSCs and progenitors.Static culture systems such as F-flasks and gaspermeableblood bags are the most widely used culture devices forexpanding hematopoietic cells.展开更多
1 IntroductionTissue engineering has emerged as a possible alter-native strategy to regenerate bone.Three components areessential:isolation and expansion of osteopogenitors ormesenchymmal stem cells,provision of appro...1 IntroductionTissue engineering has emerged as a possible alter-native strategy to regenerate bone.Three components areessential:isolation and expansion of osteopogenitors ormesenchymmal stem cells,provision of appropriate osteoin-ductive factors and an appropriately designed scaffold thatmimics the structural environment to promote bone regen-eration[1].展开更多
Introduction1Cryopreservation and prolonged storage of living cell sand tissue fragments have been an increasing interest inbiomedical engineering area during the past few decades,for the puposes of diagnostics,therap...Introduction1Cryopreservation and prolonged storage of living cell sand tissue fragments have been an increasing interest inbiomedical engineering area during the past few decades,for the puposes of diagnostics,therapy as well as funda-mental research.It will facilitate the clinic therapy andlessen the patients'pain if the autograft and heterograft ofbone used for large bulk bone loss can be successfullycryopreserved.So,the cryopreservation of osseous tissuehas a promising future and a comprehensive practical val-ue.展开更多
基金This work was supported by grants from the Int. Cooperation Project for National & Abroad Lab. of the National Natural Sciences Foundation of China (No. 2002008)The Science & Technology Foundation of Liaoning Province (No. 20022140).
文摘Objective To study large-scale expansion of SD （Sprague-Dawley） rat＇s osteoblasts in suspension culture in a rotating wall vessel bioreactor （RWVB）. Methods The bioreactor rotation speeds were adjusted in the range of 0 to 20 rpm, which could provide low shear on the rnicrocarriers around 1 dyn/cm^2. The cells were isolated via sequential digestions of neonatal （less than 3 days old） SD rat calvaria. After the primary culture and several passages, the cells were seeded onto the microcarriers and cultivated in T-flask, spinner flask and RWVB respectively. During the culture period, the cells were counted and observed under the inverted microscope for morphology every 12 h. After 7 days, the cells were evaluated with scanning electron microscope （SEM） for histological examination of the aggregates. Also, the hematoxylin-eosin （HE） staining and alkaline phosphatase （ALP） staining were performed. Moreover, von-Kossa staining and Alizarin Red S staining were carded out for mineralized nodule formation. Results The results showed that in RWVB, the cells could be expanded by more than ten times and they presented better morphology and vitality and stronger ability to form bones. Conclusions The developed RWVB can provide the culture environment with a relatively low shear force and necessary three-dimensional （3D） interactions among cells and is suitable for osteopath expansion in vitro.
基金This work was supported by Bureau of International Cooperation, NSF of China (2002008) and Science and Technology Funds of Liaoning Province (20022140).
文摘Objective To analyze the forces of rotational wall vessel （RWV） bioreactor on small tissue pieces or microcarrier particles and to determine the tracks of microcarrier particles in RWV bioreactor. Methods The motion of the microcarrier in the rotating wall vessel （RWV） bioreactor with both the inner and outer cylinders rotating was modeled by numerical simulation. Results The continuous trajectory of microcarrier particles, including the possible collision with the wall was obtained. An expression between the minimum rotational speed difference of the inner and outer cylinders and the microcarrier particle or aggregate radius could avoid collisions with either wall. The range of microcarrier radius or tissue size, which could be safely cultured in the RWV bioreactor, in terms of shear stress level, was determined. Conclusion The model works well in describing the trajectory of a heavier microcarrier particle in rotating wall vessel.
文摘1 IntroductionRecent research shows that neural stem cells mayplay an important role in the nerve injury reparation andnerve disease treatment.The shortage of the source andthe number of NSCs,however,is the main challenge forits clinic application.In thi s situation,expansion of NSCsin large scale and culture in three dimensional enviorrment are very worth of exploration.Notablely,the shearstress existed in bioreactors can cause serious cell injuryespecially for the shear sensitive cells like NSCs.
文摘1 IntroductionAnogeneic bone marrow transplantation is used forthe treatment of a variety of malignant and mnmalignanthematological disorders.Human umbilical cord blood isanother source of hematopoietic stem cells.It is acceptedthat there are definite advantages in the use of cord blood(CB)as a source of haematqpoietic progenitor cells(HPC)in selected patients with malignant and mr malig-nant haematological conditions.
文摘Introduction1Vitrification is an effective method for cryopreservation of cell s[1.2].Hbwever,cells are usually damageddue to the osmotic injuy caused by the higher concentra-tions of cryoprotective agents(CPA)during CPA remov-ing.The ice recrystallization in thawing solution can alsohurt cells serio usly.Antifreeze glycoprotein(AFGPs)isextremely efficient at inhibiting ice recrystallization[3].
文摘1 IntroductionClinical trials have demonstrated that ex vivo ex-panded hematopoietic stem cells(HSCs)and progenitorsoffer great pomise in reconstituting in vivo hematopoiesisin patients who lave undergpne intensive chemmtherapy.It is therefore necessary to develop a clinical scale culturesystem to provide the expanded HSCs and progenitors.Static culture systems such as F-flasks and gaspermeableblood bags are the most widely used culture devices forexpanding hematopoietic cells.
基金This work was supported by grantsfromThe Int.Cooperation Projectfor National&Abroad Lab.of the National Natural Sciences Foundation ofChina(2002008)The Science&Technology Foundation of Liaoningprovince(20022140)
文摘1 IntroductionTissue engineering has emerged as a possible alter-native strategy to regenerate bone.Three components areessential:isolation and expansion of osteopogenitors ormesenchymmal stem cells,provision of appropriate osteoin-ductive factors and an appropriately designed scaffold thatmimics the structural environment to promote bone regen-eration[1].
文摘Introduction1Cryopreservation and prolonged storage of living cell sand tissue fragments have been an increasing interest inbiomedical engineering area during the past few decades,for the puposes of diagnostics,therapy as well as funda-mental research.It will facilitate the clinic therapy andlessen the patients'pain if the autograft and heterograft ofbone used for large bulk bone loss can be successfullycryopreserved.So,the cryopreservation of osseous tissuehas a promising future and a comprehensive practical val-ue.
