Rapid and robust derivation of mesenchymal stem cells from human pluripotent stem cells via temporal induction of neuralized ectoderm

Abstract

Background Mesenchymal stem cells (MSCs) are emerging as the mainstay of regenerative medicine because of their ability to differentiate into multiple cell lineages. The infinite proliferative potential of human pluripotent stem cells (PSCs) grants an unlimited supply of MSCs. Despite their great potential in therapeutic applications, several drawbacks have hindered its clinical translation, including limited number of replication, compromised potential and altered function in late passages. The aim of this study is to establish an efficient method for the production of MSCs from pluripotent stem cells for potential clinical application in rare human disease Hutchinson-Gilford progeria syndrome. Results We established a robust method allowing rapid derivation of MSCs from both human iPSCs and ESCs via a temporal induction of neural ectoderm in chemically defined media. The iPSC- and ESC-derived MSCs satisfy the standard criteria of surface markers. They exhibited a high tri-lineage differentiation potential with over 90% transcriptional similarity to the primary MSCs derived from bone marrow. To evaluate the potential application of this method in disease modeling, MSCs were generated from iPSCs derived from a patient with Hutchinson-Gilford progeria syndrome (HGPS-MSCs) and from mutation-rectified HGPS-iPSCs (cHGPS-MSCs). HGPS-MSCs manifested accelerated senescence whereas mutation rectification rescued cellular senescence in HGPS-MSCs. Conclusions The robust method of MSC derivation from ESCs and iPSCs provides an efficient approach to rapidly generate sufficient MSCs for in vitro disease modeling and clinical applications.

Mesenchymal stem cells derived from CHIR99021 and TGF‑β induction remained on the colicomentum and improved cardiac function of a rat model of acute myocardium infarction

“…The induction of osteogenic, chondrogenic and adipocytic differentiation and subsequent evaluation of the induced cells was conducted as previously described ( 33 ). Osteogenic differentiation was initiated by culturing cells with osteo-inductive medium [DMEM basal medium supplemented with 10% FBS (HyClone; Cytiva), 10 mM glycerol phosphate, 50 µM ascorbate phosphate, 10 -7 M dexamethasone (MilliporeSigma; Merck KGaA) and 100 ng/ml recombinant human bone morphogenic protein-2 (Invitrogen; Thermo Fisher Scientific, Inc.)] for 2 weeks.…”

“…Cells were treated with the following different MSC differentiation mediums for 6 days: i) DMEM basal medium supplemented with 10% knockout serum replacement (Invitrogen; Thermo Fisher Scientific, Inc.), 1 mM L-glutamine, 10 mM non-essential amino acids, 50 U/ml penicillin/streptomycin (Invitrogen; Thermo Fisher Scientific, Inc.) and 0.5 µl/ml DMSO (MilliporeSigma; Merck KGaA); ii) DMEM containing 5 or 10 µM CHIR99021 (Stemgent, Inc.; REPROCELL), iii) DMEM containing 10 or 20 µg/ml TGF-β (Creative BioMart); or iv) DMEM medium containing 5 µM CHIR99021 and 10 µg/ml TGF-β. The selected concentrations of CHIR99021 and TGF-β used in the present study were selected on the basis of previous reports (20,(30)(31)(32)(33) and the induction medium was changed every other day.…”

Research progress of autoimmune diseases based on induced pluripotent stem cells

“…MSCs can be originated from bone marrow, adipose tissue, umbilical cord, and iPSCs. iPSCs-derived MSCs have almost identical phenotypes and tri-lineage differentiation ability with adipose tissue and bone marrow-derived MSCs ( Table 3 ) ( 91 , 92 ) and share approximately 90% of their transcriptome with primary MSCs derived from bone marrow ( 96 ). However, the preclinical studies on the therapeutic effects of iPSCs-derived MSCs have shown conflicting results in the animal model of IBD.…”

“…This could be because iPSCs-derived MSCs have more immature subpopulations of cells compared with tissue-derived MSCs, which experience a pluripotent state ( 92 ). In fact, we should consider aging or late passages could reduce proliferative and migration capacity, differentiation potential, and anti-inflammatory function ( 96 , 98 ), but the authors did not report the iPSCs-derived MSCs passages in the experiment ( 92 ).…”