Study Of Human Feeder Cells For Human Embryonic Stem Cells

Objective:We have comparatively evaluated a panel of six different human adult,fetal and children feeders for hESCs and have ranked them as supportive and non-supportive.To study human feeder cells differ in their capacities to support the growth of undifferentiated human embryonic stem cells, To evaluate the relationship bFGF of the expression and secretion of six lines of human fibroblasts and one line of mouse embryonic fibroblasts and growth of undifferentiated human embryonic stem cells throughout a standard hESCs culture procedure.Methods:1 Various human tissues cells were split by using trypsin-EDTA and tissue anchorage-depended.2 Seeking for the best conditions that support the hESCs from the density of feeders and the time of mitomycin acting on the Feeders. The hESCs were transferred to various feeders, and then compared the characteristics of them.3 Detecting the bFGF level secreted by various feeders using ELASE, including foreskin, testicle tissue, embryo tissue, fetal skin, fetal muscle, fetal lung tissues. The data were analyzed by SPSS 13.0 for windows. Comparison of the averages in the all samples were performed One-Way ANOVA.Relation between variables were performed by correlativity analysis(a=0.05).Result:1 All of the tested feeders support the hESCs growth for 3 passages in culture. On the basis of characteristics of the hESCs from six tissues, the order of feeders is foreskin-fetal lung-fetal muscle-fetal skin-testicle tissue-embryo tissue.2 We found that human feeder cells secreted larger quantities of bFGF than mouse feeder cells. bFGF could not be detected in culture media from mouse feeder cells. The mean of bFGF that secret by various feeders is significant at the 0.05 level between the embryo tissue group and other groups.Conclution:1 Two cell lines have been derived respectively from four tissues, including foreskin, testicle tissue, embryo tissue, fetal lung fibroblasts.We cultured successfully one cell line of fetal skin, fetal muscle fibroblasts. A lot of various cells were frozen preparing for transferring the hESCs.2 All of the tested feeders can support the hECSs growth for over 3 passages in culture, but the capability of them are not as good as the ones that growing on MEF.3 The ability of feeders layer to promote the undifferentiated growth of hESCs is attributable to its characteristic growth factor production. The relation of bFGF and growth of undifferentiated hESCs is correlation.