3 research outputs found
In Vivo Ectopic Implantation Model to Assess Human Mesenchymal Progenitor Cell Potential
Clinical interest on human mesenchymal progenitor cells (hMPC) relies on their potential applicability in cell-based therapies. An in vitro characterization is usually performed in order to define MPC potency. However, in vitro predictions not always correlate with in vivo results and thus there is no consensus in how to really assess cell potency. Our goal was to provide an in vivo testing method to define cell behavior before therapeutic usage, especially for bone tissue engineering applications. In this context, we wondered whether bone marrow stromal cells (hBMSC) would proceed in an osteogenic microenvironment. Based on previous approaches, we developed a fibrin/ceramic/BMP-2/hBMSCs compound. We implanted the compound during only 2 weeks in NOD-SCID mice, either orthotopically to assess its osteoinductive property or subcutaneously to analyze its adequacy as a cell potency testing method. Using fluorescent cell labeling and immunohistochemistry techniques, we could ascertain cell differentiation to bone, bone marrow, cartilage, adipocyte and fibrous tissue. We observed differences in cell potential among different batches of hBMSCs, which did not strictly correlate with in vitro analyses. Our data indicate that the method we have developed is reliable, rapid and reproducible to define cell potency, and may be useful for testing cells destined to bone tissue engineering purposes. Additionally, results obtained with hMPCs from other sources indicate that our method is suitable for testing any potentially implantable mesenchymal cell. Finally, we propose that this model could successfully be employed for bone marrow niche and bone tumor studies. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s12015-013-9464-1) contains supplementary material, which is available to authorized users
In vivo ectopic implantation model to assess human mesenchymal progenitor cell potential
Clinical interest on human mesenchymal progenitor cells (hMPC) relies on their potential applicability in cell-based therapies. An in vitro characterization is usually performed in order to define MPC potency. However, in vitro predictions not always correlate with in vivo results and thus there is no consensus in how to really assess cell potency. Our goal was to provide an in vivo testing method to define cell behavior before therapeutic usage, especially for bone tissue engineering applications. In this context, we wondered whether bone marrow stromal cells (hBMSC) would proceed in an osteogenic microenvironment. Based on previous approaches, we developed a fibrin/ceramic/BMP-2/hBMSCs compound. We implanted the compound during only 2 weeks in NOD-SCID mice, either orthotopically to assess its osteoinductive property or subcutaneously to analyze its adequacy as a cell potency testing method. Using fluorescent cell labeling and immunohistochemistry techniques, we could ascertain cell differentiation to bone, bone marrow, cartilage, adipocyte and fibrous tissue. We observed differences in cell potential among different batches of hBMSCs, which did not strictly correlate with in vitro analyses. Our data indicate that the method we have developed is reliable, rapid and reproducible to define cell potency, and may be useful for testing cells destined to bone tissue engineering purposes. Additionally, results obtained with hMPCs from other sources indicate that our method is suitable for testing any potentially implantable mesenchymal cell. Finally, we propose that this model could successfully be employed for bone marrow niche and bone tumor studies.This work was supported by Seventh Framework Programme of the European Commission (EuroNanomed ERA-NET initiative, REBONE project; PI10/02985FIS to J.G-C), Madrid regional government (CellCAM; P2010/BMD-2420 to J.G-C), Ministerio de Ciencia e Innovación (PI11/00377FIS to J.G-C, CSD2009-00088 to J.L.L-L.) and Mapfre foundation (SA/12/AYU/197 to Ar.A). We are grateful to Elena Andrés, David Castejón and María Encarnación Fernandez-Valle for their support in image techniques, to Daniel Baizan for his technical support in animal care, to Isabel Mirones for her technical support in immunohistochemistry and to Isabel Colmenero, pathologist, for her support in sample characterizations.S
Additional file 1: Figure S1. of Restrained Th17 response and myeloid cell infiltration into the central nervous system by human decidua-derived mesenchymal stem cells during experimental autoimmune encephalomyelitis
Foxp3 expression by T cells from untreated or DMSC-treated EAE mice (n = 9/group). Spleen cells from EAE animals were obtained at day 10 p.i. Total RNA samples were obtained from purified CD4+ cells and used to quantify Foxp3 mRNA by RT-qPCR (A). Total spleen populations from individual mice were analyzed by cytometry analysis of surface and intracellular staining with anti-CD4-APC and anti-Foxp3-PE, respectively (B). Percentages of CD4+Foxp3+ cells are shown by representative flow dot plots and by the average of the values obtained for each individual mouse from each group. Standard error of the means are shown. (PDF 150 kb