In vitro culture of multipotent adult progenitor cells purified from human bone marrow by magnetic-activated cell sorting

Abstract

Abstract: Objective To establish a method for purifying and culturing human multipotent adult progenitor cells (MAPCs) in vitro.Methods Mononuclear cells were separated from the bone marrow of healthy adult volunteers by density gradient centrifugation and cultivated in adherent culture. The plastic-adherent cultured bone marrow cells were isolated by magnetic-activated cell sorting with CD45 and GlyA magnetic microbeads, and the purity of CD45^-/GlyA^- cells evaluated by flow cytometry. Phase-contrast microscopy was used to detect morphological changes of the cells in different stages of culture.Results Approximately (5-10)×10⁴/ml MAPCs could be separated from every 1×10⁶/ml bone marrow mononuclear cells by magnetic- activated cell sorting. The viability of the cells before and after separation was (96.7±1.7)% and (96.0±2.4)%, respectively. The isolated MAPCs grew well in a self-prepared culture medium till the16th passage. The purity of CD45-/GlyA-cells separated from the bone marrow was more than 98% as examined by flow cytomety even till the 12th passage.Conclusions MAPCs derived from adult human bone marrow can be purified by magnetic-activated cell sorting with CD45 and GlyA microbeads and retain the undifferentiated state for a long time. The self-prepared culture medium is appropriate for MAPCs cultures in vitro.

CLC Number:

R318.14

MU Ning, GAO Yi, TANG Li-jun. In vitro culture of multipotent adult progenitor cells purified from human bone marrow by magnetic-activated cell sorting[J]. Journal of Southern Medical University, 2005, 25(05): 535-537.

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