文献类型：期刊文献

英文题名：Expansion of mouse sertoli cells on microcarriers

作者：Shi, B.[1];Wang, Y.[1];Zhuang, Y.[1];Chu, J.[1];Zhang, S.[1,1];Shi, X.[2];Bi, J.[3];Guo, M.[1]

机构：[1]E China Univ Chem Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Capital Med Univ, Reprod Med Ctr, Beijing, Peoples R China;[3]Univ Adelaide, Sch Chem Engn, Adelaide, SA, Australia

年份：2010

卷号：43

期号：3

起止页码：275

外文期刊名：CELL PROLIFERATION

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000277164400008)】；

基金：This study was supported by the National High Technology Research & Development Program (863 Program) of China (No. 2007AA02Z216), the National Special Fund for State Key Laboratory of Bioreactor Engineering (No. 2060204) and National Basic Research Program of China (No. 2007CB714303).

语种：英文

摘要：Background: Sertoli cells (SCs) have been described as the 'nurse cells' of the testis whose primary function is to provide essential growth factors and create an appropriate environment for development of other cells [for example, germinal and nerve stem cells (NSCs), used here]. However, the greatest challenge at present is that it is difficult to obtain sufficient SCs of normal physiological function for cell transplantation and biological medicine, largely due to traditional static culture parameter difficult to be monitored and scaled up. Objective: Operational stirred culture conditions for in vitro expansion and differentiation of SCs need to be optimized for large-scale culture. Materials and methods: In this study, the culturing process for primary SC expansion and maintaining lack of differentiation was optimized for the first time, by using microcarrier bead technology in spinner flask culture. Effects of various feeding/refreshing regimes, stirring speeds, seed inoculum levels of SCs, and concentrations of microcarrier used for expansion of mouse SCs were also explored. In addition, pH, osmotic pressure and metabolic variables including consumption rates of glucose, glutamine, amino acids, and formation rates of lactic acid and ammonia, were investigated in culture. Results: After 6 days, maximal cell densities achieved were 4.6 x 106 cells/ml for Cytodex-1 in DMEM/FBS compared to 4.8 x 105 cells/ml in static culture. Improved expansion was achieved using an inoculum of 1 x 105 cells/ml and microcarrier concentration of 3 mg/ml at stirring speed of 30 rpm. Results indicated that medium replacement (50% changed everyday) resulted in supply of nutrients and removal of waste products inhibiting cell growth, that lead to maintenance of cultures in steady state for several days. These conditions favoured preservation of SCs in the undifferentiated state and significantly increased their physiological activity and trophic function, which were assessed by co-culturing with NSCs and immunostaining. Conclusion: Data obtained in this study demonstrate the vast potential of this stirred culture system for efficient, reproducible and cost-effective expansion of SCs in vitro. The system has advantages over static culture, which has major obstacles such as lower cell density, is time-consuming and susceptible to contamination.