Functional Mesenchymal Stem Cells Derived From Human Induced Pluripotent Stem Cells Attenuate Limb Ischemia in Mice

Abstract

Background— Aging and aging-related disorders impair the survival and differentiation potential of bone marrow mesenchymal stem cells (MSCs) and limit their therapeutic efficacy. Induced pluripotent stem cells (iPSCs) may provide an alternative source of functional MSCs for tissue repair. This study aimed to generate and characterize human iPSC-derived MSCs and to investigate their biological function for the treatment of limb ischemia. Methods and Results— Human iPSCs were induced to MSC differentiation with a clinically compliant protocol. Three monoclonal, karyotypically stable, and functional MSC-like cultures were successfully isolated using a combination of CD24 − and CD105 + sorting. They did not express pluripotent-associated markers but displayed MSC surface antigens and differentiated into adipocytes, osteocytes, and chondrocytes. Transplanting iPSC-MSCs into mice significantly attenuated severe hind-limb ischemia and promoted vascular and muscle regeneration. The benefits of iPSC-MSCs on limb ischemia were superior to those of adult bone marrow MSCs. The greater potential of iPSC-MSCs may be attributable to their superior survival and engraftment after transplantation to induce vascular and muscle regeneration via direct de novo differentiation and paracrine mechanisms. Conclusions— Functional MSCs can be clonally generated, beginning at a single-cell level, from human iPSCs. Patient-specific iPSC-MSCs can be prepared as an “off-the-shelf” format for the treatment of tissue ischemia.

iPSC‐derived mesenchymal stem cells exert SCF‐dependent recovery of cigarette smoke‐induced apoptosis/proliferation imbalance in airway cells

“…First, iPSC‐MSCs have a greater expansion capacity, maintaining differentiation potential and normal karyotypes for 120 doublings, while BM‐MSCs get senescent after 20 doublings 11. Second, BM‐MSCs may present reduced survival and differentiation ability when isolated from aged subject or patients with ageing‐related disorders 12, 13, 14, 15.…”

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

“…iPSC-MSCs could form mature mineralized structures that were histologically similar to mature bone, facilitating periodontal regeneration [39,40]. Transplanting iPSC-MSCs attenuated severe hindlimb ischemia and improved the hepatic function in mouse models [33,41,42]. These results suggested that iPSC-MSCs have high potential for tissue-engineering applications.…”

“…The iPSC lines were prepared from iPSCs, which were reprogrammed from human fibroblast cells, and differentiated into MSCs according to a previously described protocol [33]. Briefly, MSCs were purified by sorting for CD105 + /CD24 -cells and maintained in a medium containing 90% knockout Dulbecco's Modified Eagle's Medium (Gibco, Invitrogen Corporation, Carlsbad, CA) supplemented with 10% serum replacement medium (Gibco) and basic fibroblast growth factor (10 ng/mL; Gibco).…”

“…Briefly, MSCs were purified by sorting for CD105 + /CD24 -cells and maintained in a medium containing 90% knockout Dulbecco's Modified Eagle's Medium (Gibco, Invitrogen Corporation, Carlsbad, CA) supplemented with 10% serum replacement medium (Gibco) and basic fibroblast growth factor (10 ng/mL; Gibco). The morphology of iPSC-MSCs was very similar to BMMSCs and they have the capacity to differentiate into osteoblasts, adipocytes, and chondroblasts [33]. Millipore's FlowCellectÔ Human Mesenchymal Stem Cell Characterization Kit was used for the phenotypic analysis of iPSCMSCs.…”

“…Therefore, it is important to identify alternative sources of MSCs before they can be used as a mainstream treatment for organ transplantation. A breakthrough in the generation of humaninduced pluripotent stem cells (iPSCs) from adult somatic cells offered the possibility of generating a high yield of MSCs [33][34][35].…”