ESTABLISHMENT OF CLONAL MARROW STEM CELL LINES FROM RAT OR RABBIT BONE MARROW USING SUBFRACTIONATION CULTURING METHOD

초록

Bone marrow has been considered to contain many different types of progenitor or stem cells. We previously reported a new strategy that provides for the rapid establishment of human clonal marrow stromal/stem cell (hcMSC) lines with a relatively small amount of bone marrow aspirate (Song et al., 2008). The rationale of this method, called subfractionation culturing method (SCM), is that different types of marrow stromal or stem cells can be separated based on their natural cell densities by a series of temporal transfers of the culture supernatant to subsequent dishes where the low density stem cells with high proliferation and differentiation potential may be fractionated. Several different clonal marrow stromal/stem cell lines were established with rat or rabbit bone marrow aspirate and characterized for their cell surface epitopes, cell proliferation , differentiation potential, and lineage-specific gene expression. Five different rat or rabbit clonal marrow stromal/stem cell lines, rcMSC or rbcMSC respectively, were established with bone marrow of Sprague Dawley rat or New Zealand white rabbit. The overall profiles of the cell surface epitopes of the newly established rat or rabbit cMSC lines were similar to those of the known rat or rabbit MSCs. However, each rat or rabbit cMSC lines exhibited some variation in the expression of some epitopes, for example, CD31, CD44, and CD 106 in rat or CD90 and CD117 in rabbit. These cMSC lines showed different expansion potential and were capable of differentiating into mesodermal lineages with some differences in differentiation capability. These data suggest that the newly designed protocol, subfractionation culturing method, is an efficient method to establish single-cell derived cMSC lines with rat or rabbit bone marrow and some of these rat or rabbit cMSC lines possess stem cell characteristics and exhibit differences in cell surface epitopes, expansion capability, differentiation potential and lineage-specific gene ex