[Effect of monoclonal or different cell seeding densities on the differentiation potential of immortalized human mesenchymal stem cells in vitro and in vivo]

Abstract

Objective: To determine whether monoclonality or different cell seeding densities could influence the differentiation potential of immortalized human mesenchymal stem cells (hMSC-TERT) and to find an effective cultural method of hMSC-TERT in vitro.

Methods: From the parental hMSC-TERT cell line, we derived 30 monoclonal cell lines and two independent cell lines based on different plating densities during expansion in culture. Their adipocytic, osteogenic, neuronal differentiation potential in vitro and multidirectional differentiation potential in vivo were analyzed by immunohistochemistry for pathologic tissue markers.

Results: Monoclonal cell lines and the cell line derived at low seeding density had a lower differentiation potential in vitro than the cell line derived at higher cell seeding density. The differentiation potential of monoclonal hMSC-TERT cells were dissimilar. Some of monoclonal hMSC-TERT lines expressed epithelial differentiation potential in vivo while the parental hMSC-TERT cells line did not.

Conclusion: Multiclonal hMSC-TERT cells cultured in high seeding density can keep the differentiation potential, cloning the hMSC-TERT cells before transplantation to find the special clones for special purpose of transplantation.