Proliferative Potential of Multipotent Mesenchymal Stromal Cells from Human Bone Marrow

Abstract

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.

Age-Related Differences in Rat Multipotent Mesenchymal Stromal Bone Marrow Cells

“…A correlation of the cell lifetime and doubling number in culture with species-specifi c lifetime was demonstrated in many types of cultured cell. A dependence of replicative activity on donor's age was shown for human hepatocytes, keratinocytes, arterial smooth muscle cells, and MMSC [1,5,6,11]. The num- ber of determined and inducible osteogenic precursors also decreased with age [11,20].…”

“…The number of hemopoietic precursors in BM during aging decreased less markedly and rapidly [12]. More intricate infl uences of the age and external and internal factors (which leads to pronounced individual differences within the same age group) on the BM pool of mesenchymal precursors were demonstrated by a group headed by N. I. Drize [5].Morphologically, aging cells look larger and express -galactosidase [14,18,20,21]. At the same time, some authors reported the absence of age-related differences.…”

Trophic Actions of Bone Marrow-Derived Mesenchymal Stromal Cells for Muscle Repair/Regeneration

“…MSCs possess many biological properties that make these cells ideal candidates for tissue engineering and regenerative medicine. These properties include: the ease of accessibility for isolation from the patients or bone marrow banks; the high expansion potential [46,47]; and, the presumptive plasticity, that is, being able to differentiate in vitro , not only into mesenchymal, but also non-mesenchymal lineages, including myoblasts [48], cardiomyocytes [49,50], hepatocyte-like cells [51], neuronal and neuroglial cells [52,53] and endothelial cells [54]. Moreover, when transplanted systemically, MSCs are able to migrate and home to the specific site of injury [55,56] and exert anti-inflammatory and immunosuppressive effects thus allowing a potential for their autologous and allogenic use [57,58,59].…”

Efficiency of Exosome Production Correlates Inversely with the Developmental Maturity of MSC Donor

“…Unlike the controversial embryonic stem cells, MSCs could be isolated from many ethically palatable tissues such as bone marrow [16,17], adipose tissue [17,18], liver [19,20], muscle [21,22], amniotic fluid [23,24], placenta [25,26], umbilical cord blood [16,27], dental pulp [28,29]. However, it is generally observed that the biological activity and therapeutic potency of MSCs correlate inversely with developmental stage of the donor [30][31][32][33][34][35][36][37][38][39][40][41][42][43][44][45][46][47][48]. Therefore, MSCs from ethically and socially controversial but "young" tissues such as fetal tissues [49] and human embryonic stem cells (ESCs) [50] continue to be investigated for their therapeutic potential.…”