Proliferative potential of multipotent mesenchymal stromal cells from human bone marrow.
We studied the capacity of multipotent mesenchymal stromal cells isolated from human bone marrow (BM) to long-term passaging, cloning, and re-cloning. Initial multipotent mesenchymal stromal cells and cells after gene labeling were studied. Multipotent mesenchymal stromal cells were obtained from donors (13-59 years) and cultured for 7 passages. Third generation lentivector was used for delivery of green fluorescent protein marker gene. The procedure of infection revealed reduced proliferative potential of multipotent mesenchymal stromal cells from elder donors. Hierarchy of precursor cells differing by their proliferative potential was demonstrated in the culture of multipotent mesenchymal stromal cells. Three categories of multipotent mesenchymal stromal cells were identified: mature cells incapable of proliferation (75.7±2.4% population) and cells with low and high proliferative potential (17.6±2.1 and 6.7±0.3%, respectively). The relative content of these cells insignificantly differed from passage to passage. The efficiency of cloning also remains stable, but re-cloning capacity sharply decreased after passage 3 and completely disappeared in multipotent mesenchymal stromal cells after cryopreservation. Thus, cultured multipotent mesenchymal stromal cells represent a heterogeneous and hierarchically organized population and the characteristics of this population depend of the duration of culturing and age of BM donor. This should be taken into account when using multipotent mesenchymal stromal cells in clinical practice.
Hematological Research Center, Ministry of Health Care and Social Development of the Russian Federation, Moscow, Russia.
O A
OA
Zhironkina
I N
IN
Shipounova
A E
AE
Bigildeev
N V
NV
Sats
N A
NA
Petinati
N I
NI
Drize
D000293:Adolescent; D000328:Adult; D000367:Age Factors; D001854:Bone Marrow Cells / Q000166:cytology* / Q000502:physiology; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation; D002478:Cells, Cultured; D002999:Clone Cells; D003114:Colony-Forming Units Assay; D015925:Cryopreservation; D005260:Female; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D016086:Lentivirus; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology; D008875:Middle Aged; D039902:Multipotent Stem Cells / Q000166:cytology* / Q000502:physiology
D049452:Green Fluorescent Proteins