Studies of growth factor on mammalian cell growth

• Main Authors: Kuo Wei Wu, 吳國維
• Other Authors: C. H. Liu
• Format: Others
• Published: 2008
• Online Access: http://ndltd.ncl.edu.tw/handle/98227645482833819768

碩士 === 長庚大學 === 生化與生醫工程研究所 === 96 === Chinese hamster ovary (CHO) cells are commonly used in the biotechnology industry to produce recombinant proteins. The development of serum-free media in order to maintain mammalian cell viability and reduce cell death in the absence of serum is highly desirable. Our aim was to find new growth factors that could maintain viability and further enhance the proliferation of CHO cells under serum-free conditions. Macrophage colony-stimulating factor (M-CSF)-secreting CHO cells were used as a model cell line in this study. The stimulatory effects of different compounds including two growth factors on the proliferation of CHO cells were evaluated. Flow cytometry and fluorescence microscopy were used to analyze the cell cycle and monitor apoptotic nuclei during the serum-free culture of CHO cells. Among the tested compounds, insulin and basic fibroblast growth factor (bFGF) each individually significantly prevented CHO cells from dying and helped maintain cell viability, but cells were unable to reenter the cell cycle and proliferate. Furthermore, simultaneous supplementation with insulin and bFGF synergistically promoted the growth of CHO cells and recombinant M-CSF synthesis in basal DMEM/F12 medium. The results presented here provide useful information for the design and development of serum-free media as well as for antiapoptotic studies of CHO cells under serum-free conditions. In preliminary results of mesenchymal stem cells (MSC), we found out the commercial serum substitute (lipumin), ITS, bFGF, insulin had the significantly effects on cell proliferation under serum-free conditions. After, we also used the commercial phenotype microarray to evaluate the effects of new serum substitutes. Finally, Our aim is to develop serum-free medium for MSC’s proliferation and differentiation.