An Alternative Method for Long-Term Culture of Chicken Embryonic Stem Cell In Vitro

Figure 1

The effect of feeders and factors on the development of avian blastodermal cells. (a) bFGF is vital to the culture of avian blastodermal cells in vitro. cESCs were plated in the presence of or in the absence of growth factors as indicated in the figure. Crystal violet staining was performed after 4 passages. (b) The areas of colonies with crystal violet staining were counted by Image-Pro Plus. Compared to the control group, there is a significant difference (). ^#Compared to the LIF, SCF, and LIF + SCF groups, there is a significant difference (). (c) The combination of 20 ng/μl each of hLIF, hbFGF, and mSCF was the optimal concentration. cESCs were plated in 0, 10, and 20 ng/μl each of hLIF, hbFGF, and mSCF, respectively. Four passages later, crystal violet staining was performed. (d) The areas of colonies were calculated by Image-Pro Plus. The colony area in the presence of 20 ng/μl growth factors was the biggest (). (e) The inactivated DF-1 feeder was the optimal feeder for supporting cESCs in vitro compared to STO and pCEF feeders. (f) The areas of colonies with crystal violet staining after 4 passages were counted by Image-Pro Plus. The colony area on the DF-1 feeder was the biggest (). (g) The cultured cESCs after 4 passages on different feeders were performed by DAPI (blue), EDU (green), and NANOG (orange) staining. Scale bar: 150 μm. (h) The bars indicated the positive proliferation rate of the cultured cESCs after 4 passages on different feeders. Cells on the DF-1 feeder and pCEF feeder showed the highest proliferation rate (). All results are expressed as the mean ± SD of three independent experiments. Scale bar: 100 μm.

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