[Cultivation and purification of high proliferative potential endothelial progenitor cells from murine bone marrow]

The present study was designed to culture and purify high proliferative potential endothelial progenitor cells (HPP-EPCs) from murine bone marrow and their progeny cells using a culture system which has been established by us recently, in order for more intensive researches on these cells. Fresh murine bone marrow cells were preplated to allow the preferential attachment of non-EPCs to tissue culture plates and then unattached cells were repreplated in the culture system containing the bone marrow endothelial cell line-conditioned medium (BMEC-CM). The colonies containing about 2 x 10(4) cells were collected respectively and single colony-derived cells were cultured for their expansion in the above-mentioned culture system. These single colonies were able to yield 8 x 10(6) progeny cells. The endothelial-lineage characteristics of expanded cells were confirmed by immunofluorescent staining for CD31 and vWF, FITC-UEA-1 binding and Dil-Ac-LDL uptake. These data suggest that murine bone marrow HPP-EPCs can proliferate exuberantly so that their progeny cells can be obtained with high yield by using the single colony-derived cell culture in the presence of BMEC-CM.