The comparison of both protein spots and mRNA levels between T3 MEF and T3 CMMEF cells exhibited the most similarity, while that of T3 HDF and T3 MEF cells had lowest similarity. Discussion The hES T3 cell line with normal female karyotype, one of five hES cell lines derived in our laboratory, was used to differentiate into autogeneic www.selleckchem.com/products/AG-014699.html fibroblast like cells as feeder to support the undifferentiated growth of hES T3 cells for 14 passages according to the previously published procedure. Stojkovic et al. reported that the hES cells cultured on autogeneic feeder and Matrigel in the presence of autogeneic conditioned medium for 44 and 14 passages, respectively, still main tained normal karyotype and expressed hES markers such as TRA 1 60, SSEA 4 and GTCM 2.

This autogeneic fee der system was further shown to permit continuous growth of pluripotent hES cells as demonstrated by the formation of teratoma in SCID mice and in vitro differen tiation. In this investigation, a feeder free culture on Matrigel in medium conditioned by these autogeneic fee der cells was established to maintain the undif ferentiated growth of hES T3 cells for 8 passages. The gene expression profiles of mRNAs, miR NAs and GSK-3 proteins among the undifferentiated T3 HDF, T3 CMHDF, T3 MEF and T3 CMMEF cells were shown to be very similar. In recent years, many improvements on standard MEF culture have been reported to develop xeno free culture systems of hES cells for future clinical applications. To our knowledge, this investigation is the first report that systematically compared and demonstrated the similar expression profiles of mRNAs, miRNAs and proteins among different feeders and condi tioned media.

However, many more passages of the undifferentiated growth of hES T3 cells on autogeneic T3HDF feeder and feeder free on Matrigel in the T3HDF conditioned medium should be carried out and their dif ferentiation capacities should also be demonstrated using formation of embryoid bodies in vitro and or teratoma in SCID mice in the future investigation. The abundantly expressed genes of T3 HDF, T3 CMHD, T3 MEF and T3 CMMEF cells were found to play prominent roles in signaling pathways and GO pro cess networks. Three of the top 10 GeneGo canonical pathway maps and four of the top 10 GO process net works of the common and or similar genes among these four cell populations were involved in development.

Their number 1 pathway was the role of Activin A in cell differentiation and proliferation, and the importance of first Activin Nodal TGFb family members in the maintenance of pluripotency of hES cells is widely established. Among these common and or similar genes, cell adhe sion was also involved in three of the top 10 GeneGo canonical pathway maps and two of the top 10 GO pro cess networks.