Investigation of cocktails of growth factors for proliferative zonal growth plate using multi-array microfluidic platform
Yeo, Qiu Ju
Date of Issue2015
School of Chemical and Biomedical Engineering
A major problem in the development of engineering an in-vitro growth plate is the premature entrance of the chondrocytes to the hypertrophic stage without the production of Collagen type II (Col2). The study explored the use of a cocktail of growth factors, in particularly, TGF-β3, FGF-2 and insulin, to differentiate hMSCs into and maintain the chondrocytes at proliferative zone of growth plate with the use of microfluidic platform. TGF-β3 was added in all samples for chondrogenic differentiation in hMSCs. FGF-2 and insulin, on the other hand, were added at different time points to investigate the best condition to aid the induction of proliferative chondrogenesis, at the same time inhibit hypertrophy. Negative alcian blue and lack of significant difference in proliferation rates between cells cultured with TGF-β3 only (control group) and other growth factors cocktails suggest that the cells were still in early proliferative stage after Day 14. Hence, the experiment was then extended to 4-week. The proliferation rates were shown to be significantly higher than control group when FGF-2 was added from Day 0. Unexpectedly, cells feeded with FGF-2 in combination with insulin, did not exhibit difference in proliferation rates compared to supplement of FGF-2 alone. Alcian blue stains also showed some proteoglycan detected, but not abundant. It was thought that optimal cell density was not reached for proliferation due to frequent evaporation of medium as a result of the weak bonding between the PDMS chip and base. Further analysis and studies are still required to identify the chondrogenic stage of the cells and determine the progress of the investigation.
Final Year Project (FYP)
Nanyang Technological University