微载体浓度与细胞接种密度对ST细胞生长的影响  

The Effects of Microcarrier Concentration and Cell Density on the Growth of Swine Testicle Cells

作　　者：陈以衡[1] 陶姝宇[1] 刘旭平[1] 谭文松[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

出　　处：《生物技术通报》2016年第4期242-250,共9页Biotechnology Bulletin

基　　金：国家自然科学基金项目(21206040;21406066);国家"863"计划项目(2012AA02A303);国家重大专项(2013ZX10004003-003-003);中央高校基本科研业务费(WF1214035)

摘　　要：选择合适的微载体浓度、细胞接种密度以提高微载体利用率,优化微载体培养体系猪睾丸细胞(Swine testicle cells)的贴附生长与维持。使用DMEM补加10%血清、LSM(Low serum medium)两种培养基考察微载体浓度、细胞接种密度对细胞生长维持的影响,进而比较ST细胞在不同条件下对Cytodex1微载体的利用率。结果显示,使用LSM在T150方瓶中连续传代培养30d,平均比生长速率为0.626d^(0-1),是DMEM补加10%FBS培养基的1.15倍。选择10×10~5cells/mL细胞接种3g/LCytodex1搅拌瓶体系,最大细胞密度为38.3×10~5cells/mL,微载体利用率上升到58.8%。在灌注培养体系中培养ST细胞15d,最终细胞密度达到36.6×10~5cells/mL,扩增了13.6倍。微载体悬浮培养的使用一方面有利于ST细胞的贴附与生长,实现高密度生长,另一方面增加了微载体的使用成本,选择合适的微载体浓度、细胞接种密度,能够最大化利用微载体与培养基中的营养物质实现细胞的最优生长。This study is to select the suitable microcarrier concentration and cell seeding density for enhancing the utilization of microcarrier, and optimize the adhered growth and maintenance of ST （ swine Testicle ） cells. The effects of microcarrier concentration and seeding density on cell. growth and maintenance were investigated using 2 different mediums ： DMEM supplemented 10% serum or low serum medium （ LSM ） . Further, the utilizations of Cytodex 1 microearrier by ST cells under different conditions were compared. Results were as below：continuous culture of ST cells in T150 flask using LSM lasted for 30 days, and the average specific growth rate was 0.626 d-~, which was 1.15 times of that using DMEM supplemented with 10% FBS. The maximum cell density was 38.3 x 10 cells/mL and the utilization was up to 58.8% when microcarrier concentration was 3 g/L Cytodex 1 in mixing bottle and seeding density was 10 x l0s cells/mL. Furthermore, when cultured in perfusion system for 15 days, the final ST cell density reached 36.6 x l0s cells/mL, which amplified 14.6 times. Conclusively, the uses of serum and microcarrier are favorable for ST cell adhesion and growth to achieve high cell density, but also this increases the cost of introducing microcarrier. Maximum utilization of microcarrier and nutrients in the medium, and the optimal growth and maintenance could be achieved through selecting suitable microcarrier concentration and seeding density. The results in this work provide guidance for further industrial-scale microcarrier culture system producing CSFV vaccine.

关 键 词：微载体 ST细胞 猪瘟病毒 细胞密度 

分 类 号：Q813[生物学—生物工程]