Characterization of Adult Stem/Progenitor Cell Populations from Bone Marrow in a Three-Dimensional Collagen Gel Culture System

Abstract

Stem cell transplantation therapy using mesenchymal stem cells (MSCs) is considered a useful strategy. Although MSCs are commonly isolated by exploiting their plastic adherence, several studies have suggested that there are other populations of stem and/or osteoprogenitor cells that are removed from primary culture during media replacement. Therefore, we developed a three-dimensional (3D) culture system in which adherent and nonadherent stem cells are selected and expanded. Here, we described the characterization of 3D culturederived cell populations in vitro and the capacity of these cells to differentiate into bone and/or cartilage tissue when placed inside of demineralized bone matrix (DBM) cylinders, implanted subcutaneously into the backs of rat for 2, 4, and 8 weeks. Our results demonstrates that 3D culture cells were a heterogeneous population of uncommitted cells that express pluripotent-, hematopoietic-, mesenchymal-, and endothelial-specific markers in vitro and can undergo osteogenic differentiation in vivo.

A Novel Human TGF-β1 Fusion Protein in Combination with rhBMP-2 Increases Chondro-Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells

“…Results showed that Nanog, Oct4 and Sox2 were significantly highly expressed in 3D culture at days 10 and 14 but was absent in ADH cells. These findings are also in agreement with those previously reported by our group [36]. Nanog, Oct4 and Sox2 target genes, identified in embryonic stem cells (ESCs), are known to overlap substantially, suggesting they collaborate to regulate a common set of genes governing pluripotency, self-renewal, and cell fate determination [47,48,49,50,51].…”

“…3D culture revealed a heterogeneous population of small round cells [36,37]. These results were consistent with our previous studies with human BM-derived cells and commercial rhTGF-β1 in the 3D culture [36], although the immunophenotyping was difficult because of the limited availability of antibodies that show cross-reactivity with the rat.…”

“…3D culture revealed a heterogeneous population of small round cells [36,37]. These results were consistent with our previous studies with human BM-derived cells and commercial rhTGF-β1 in the 3D culture [36], although the immunophenotyping was difficult because of the limited availability of antibodies that show cross-reactivity with the rat. The study of the surface marker profile showed positive cells for hematopoietic (CD34, CD45), mesenchymal (CD29, CD105, CD166, CD271, STRO-1) and endothelial markers (CD34, CD133) [38,39,40,41,42].…”

“…Comparing 3D cultures at days 10 and 16, we observed an increase in the expression of mesenchymal cell markers, while EPCs-like populations (CD34+/CD133+/CD45−) significantly decreased over time. Furthermore, our previous results demonstrated that EPCs-like populations decreased in favor of mature endothelial phenotypes (CD146+/KDR+) [43,44], demonstrating that our 3D culture system could be used as a strategy to promote angiogenesis and to improve bone regeneration [36,45,46]. …”

New perspectives for articular cartilage repair treatment through tissue engineering: A contemporary review

“…Increasing the source of cells for artificial repair of cartilage defects is becoming adipose tissue [27,28] , etc., so then, we can get a variable number of cells from a different tissue [29,30] . MSCs show a high proliferation and differentiation potential, although coming from different tissue, and have an uneven chondrogenic differentiation capacity probably related to the special cytokines, growth factor and induction molecules composition of the medium [31,32] . Marrow stimulating techniques attempt to solve articular cartilage damage through an arthroscopic procedure.…”