Prolonged ex vivo culture of human bone marrow mesenchymal stem cells influences their supportive activity toward NOD/SCID-repopulating cells and committed progenitor cells of B lymphoid and myeloid lineages

Abstract

The online version of this article has a supplementary appendix. BackgroundBone marrow mesenchymal stem cells support proliferation and differentiation of hematopoietic progenitor cells in vitro. Since these cells constitute a rare subset of bone marrow cells, mesenchymal stem cell preparations for clinical purposes require a preparative step of ex vivo multiplication. The aim of our study was to analyze the influence of culture duration on mesenchymal stem cell supportive activity. Design and MethodsMesenchymal stem cells were expanded for up to ten passages. These cells and CD34+ cells were seeded in cytokine-free co-cultures after which the phenotype, clonogenic capacity and in vivo repopulating activity of harvested hematopoietic cells were assessed. ResultsEarly passage mesenchymal stem cells supported hematopoietic progenitor cell expansion and differentiation toward both B lymphoid and myeloid lineages. Late passage mesenchymal stem cells did not support hematopoietic progenitor cell and myeloid cell outgrowth but maintained B-cell supportive ability. In vitro maintenance of NOD/SCID mouse repopulating cells cultured for 1 week in contact with mesenchymal stem cells was effective until the fourth passage of the mesenchymal cells and declined thereafter. The levels of engraftment of CD34 + cells in NOD/SCID mice was higher when these cells were co-injected with early passage mesenchymal stem cells; however mesenchymal cells expanded beyond nine passages were ineffective in promoting CD34 + cell engraftment. Non-contact cultures indicated that mesenchymal stem cell supportive activity involved diffusible factors. Among these, interleukins 6 and 8 contributed to the supportive activity of early passage mesenchymal stem cells but not to those of late passage cells. The phenotype, as well as fat, bone and cartilage differentiation capacity, of mesenchymal stem cells did not change during their culture. ConclusionsExtended culture of mesenchymal stem cells alters the ability of these cells to support hematopoietic progenitor cells without causing concomitant changes in their phenotype or differentiation capacity. Haematologica. 2010; 95:47-56. doi:10.3324/haematol.2009 This is an open-access paper.

iPSC-MSCs Combined with Low-Dose Rapamycin Induced Islet Allograft Tolerance Through Suppressing Th1 and Enhancing Regulatory T-Cell Differentiation

“…MSCs have previously demonstrated their capacity to facilitate regeneration and regulate immune responses in a range of animal models; however, major factors related to life span and tumorigenicity limit their widespread use in a clinical setting [28][29][30][31][32]. Recent reports have described MSC-like cells derived from iPSCs [33][34][35] with a greater proliferative capacity, lower immunogenicity, and greater immunoregulatory function compared with primary MSC cultures [33,43,45].…”

“…Successive passages slow the proliferation rate, and MSCs progressively lose their multipotency and lack immunosuppressive activity. In addition, aging and age-related disorders significantly impair the survival and differentiation potential of BM-MSCs, thus limiting their therapeutic efficacy [28][29][30][31][32]. Therefore, it is important to identify alternative sources of MSCs before they can be used as a mainstream treatment for organ transplantation.…”

Mesenchymal Stem Cells Induce Granulocytic Differentiation of Acute Promyelocytic Leukemic Cells via IL-6 and MEK/ERK Pathways

“…Recently, IL-6 was identified to mediate human BMMSCs' contribution to in vitro differentiation of hematopoietic progenitor cells toward myeloid and lymphoid cells [35]. Xie et al reported that IL-6 enhanced retinoic acid-induced granulocytic differentiation of APL cells [49].…”

“…Based on these results, we ruled out the possibility of participation of G-CSF and PGE2 in UC-MSCs' differentiation induction. Recently, IL-6 was identified to mediate human bone marrow MSCs' (BMMSCs) contribution to in vitro differentiation of hematopoietic progenitor cells toward myeloid and lymphoid cells [35]. Xie et al reported that IL-6 dramatically enhanced retinoic acid-induced granulocytic differentiation of HL-60 cells (a leukemic cell line derived from M2) [49].…”

“…Nevertheless, little is known about the influence of MSCs on the differentiation of tumor cells. Numerous reports demonstrated that MSCs enhanced the differentiation of normal hematopoietic progenitor cells toward both myeloid and lymphoid lineages [30][31][32][33][34][35]. However, whether MSCs also play a role in regulating the differentiation of leukemic stem/progenitor cells remains unknown.…”