Serum-free media studies of human embryonic stem cell culture on microcarriers

• Main Author: Chen, Xiaoli.
• Other Authors: Alastair Campbell Ritchie
• Format: Final Year Project
• Language: English
• Published: 2009
• Subjects: DRNTU::Engineering::Chemical engineering::Biotechnology
• Online Access: http://hdl.handle.net/10356/16505
• Institution: Nanyang Technological University

The 2 key properties of human embryonic stem cells (hESC), namely pluripotency and unlimited self-renewal, give them a great potential in the area of stem cell research and applications in cell therapy. It has been shown that human embryonic stem cells (hESC) can be grown and maintained on microcarriers using both defined, serum-free media, and MEF-CM. In addition, microcarrier cultures were able to produce higher cell numbers than the conventional 2D colony cultures with their pluripotent properties intact due to the availability of surface area. However, high starting glucose levels in serum-free media results in high lactate production and low pH. Cell growth kinetics, metabolic rates, controlled metabolites feeding as well as alternative carbon sources were studied. Results showed that lactate-to-glucose ratio of hESC in microcarrier cultures was within the range of 1.8-1.9, while ammonium-to-glutamine ratio of hESC was found to be approximately 0.7 or 0.8. Presence of lactate had a detrimental effect on hESC growth as reflected in a significantly lower count. Alternative sources of carbon, fructose and galactose showed to fail at maintaining hESC cultures. In addition, controlling the amount of glucose present in the media at a lower level generated a high lactate-to-glucose ratio of 2.6. This highly indicated that the cells are obtaining carbon source from elsewhere.