Abstract
Adult human bone marrow stromal cells (hBMSC) are important for many scientific purposes because of their multipotency, availability, and relatively easy handling. They are frequently used to study osteogenesis in vitro. Most commonly, hBMSC are isolated from bone marrow aspirates collected in clinical routine and cultured under the “aspect plastic adherence” without any further selection. Owing to the random donor population, they show a broad heterogeneity. Here, the osteogenic differentiation potential of 531 hBMSC was analyzed. The data were supplied to correlation analysis involving donor age, gender, and body mass index. hBMSC preparations were characterized as follows: (a) how many passages the osteogenic characteristics are stable in and (b) the influence of supplements and culture duration on osteogenic parameters (tissue nonspecific alkaline phosphatase (TNAP), octamer binding transcription factor 4, core-binding factor alpha-1, parathyroid hormone receptor, bone gla protein, and peroxisome proliferator-activated protein γ). The results show that no strong prediction could be made from donor data to the osteogenic differentiation potential; only the ratio of induced TNAP to endogenous TNAP could be a reliable criterion. The results give evidence that hBMSC cultures are stable until passage 7 without substantial loss of differentiation potential and that established differentiation protocols lead to osteoblast-like cells but not to fully authentic osteoblasts.
Highlights
Human bone marrow stromal cells are widely used for tissue engineering approaches, to evaluate the performance of diverse biomaterials, to study tissue development and regeneration, and to understand molecular issues of cell differentiation [1, 2]
The passage 1 (P1) cells were transferred into freezing medium containing 90% of human serum albumin solution and 10% of dimethylsulfoxide (Wak Chemie Medical GmbH, Steinbach, Germany) and stored in liquid nitrogen until subcultivation
Osteogenic Differentiation Potential of Human Bone Marrow Stromal Cells: Evaluation of Donor Variation. The aim of this experimental setting was to evaluate the osteogenic differentiation potential of Human bone marrow stromal cells (hBMSC) in vitro and to determine how reliably hBMSC differentiate into osteoblastlike cells
Summary
Human bone marrow stromal cells (hBMSC; usual synonyms bone marrow stem cells, mesenchymal stromal cells, and mesenchymal stem cells) are widely used for tissue engineering approaches, to evaluate the performance of diverse biomaterials, to study tissue development and regeneration, and to understand molecular issues of cell differentiation [1, 2]. Pittenger et al [3, 4] described the clonal multilineage potential of hBMSC and published first characterization criteria and differentiation protocols. HBMSC can be differentiated in vitro into cells with osteogenic, adipogenic, or chondrogenic features by adding particular supplements to the culture medium. The International Society for Cellular Therapy (ISCT) described established, clearly defined minimal characteristics of multipotent mesenchymal stromal cells: (1) adherence to cell culture plastic under standard conditions, (2) expression of certain surface markers (CD105, CD73, and CD90), (3) absence of CD45, CD34, CD11b or CD14, CD79α or CD19, and HLA-DR, and (4) trilineage differentiation capacity in vitro confirmed by staining of mineralized nodules (osteoblasts), collagen type II or glycosaminoglycans (chondrocytes), and lipid droplet accumulation (adipocytes) [5,6,7]. Beside the “minimal essential markers” propagated by ISCT, a broad panel of other markers as reviewed [9,10,11,12] was used for BMSC identification and characterization, supplemented recently by leptin receptor as a marker of
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