[In vitro culture of multipotent adult progenitor cells purified from human bone marrow by magnetic-activated cell sorting].
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)x10(4)/ml MAPCs could be separated from every 1x10(6)/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- 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.
Mu
Ning
N
Department of General Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China. mnwsq@163.com
Gao
Yi
Y
Tang
Li-jun
LJ
D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D003470:Culture Media; D006801:Humans; D017493:Leukocyte Common Antigens / Q000378:metabolism; D039902:Multipotent Stem Cells / Q000166:cytology*
D003470:Culture Media; D017493:Leukocyte Common Antigens