More Homegeneous MSCs for Clinical Application

초록

This presentation will discuss about current strategies of isolation and characterization of human marrow stromal/stem cells (MSCs). In addition, it will also introduce our newly developed protocol that provides for the rapid establishment of human clonal marrow stem cell (hcMSC) lines with a relatively small amount of bone marrow (BM) aspirate. Human cMSC lines were generated with human BM aspirates using the new protocol, called the subfractionation culturing method. The newly established hcMSC lines were analyzed for their cell surface epitopes by FACS, differentiation potential by in vitro differentiation assays, lineage specific gene expression by RT-PCR, and cytokine secretion by ELISA. The overall profile of the cell surface epitopes of the hcMSC lines was similar to those of the previously known MSCs. Some of these hcMSC lines were capable of differentiating into multi-lineages with some differences in differentiation capability. Furthermore, these hcMSC lines secrete high levels of TGF-β1, LIF, TGF-α, and IL-10, again with some variation in each cell line. The newly designed protocol may be an efficient method to isolate homogeneous populations of hcMSCs with a relatively small amount of bone marrow sample and have advantages for basic research and clinical purposes. Some of the newly established hcMSC lines possess stem cell characteristics and they exhibit some differences in cell surface epitopes, differentiation potential, lineage-specific gene expression, and cytokine secretion.