Evaluation of human induced pluripotent stem cell (ipsc)-derived mesenchymal stromal/stem cells (msc) for use in cell-based therapy

Abstract

Donor procurement makes the logistics of obtaining human bone marrow (BM) as a source of mesenchymal stem/ stromal cells (MSCs) complicated. Although BM MSCs from adult donors still have satisfactory abilities to be used for regenerative medicine purposes, from the manufacturing perspective, the best donors are in the pediatric age range. In addition, the procedure requires planning and it is painful; the derivation and identification of the MSCs according to ISCT are still controversial among scientists, and the isolation and quality of cells also varies among donors. Therefore, the MSCs therapy field is constantly looking for alternative sources.
We hypothesized that human MSC derived from induced pluripotent stem cells (iPSCs) have biological qualities of native MSC and could be scaled up and used for therapeutic purposes. To test the hypothesis, we isolated native MSC (nMSC) from umbilical cord’s Wharton’s Jelly (WJ) of two donors (#012 and #013) in xeno-free conditions. Next, we reprogrammed them into iPSC (iPSC012 and iPSC013) and subsequently differentiated them back into MSC using two different protocols ARG and TEX (iMSC012ARG, iMSC012TEX, iMSC013ARG, and iMSC013TEX). Using the same protocols, we also differentiated the clinical grade human embryonic stem cell line (ESC) KCL034 into MSC (eMSC034ARG and eMSC034TEX). To assess which of the two differentiation protocols worked better, we compared differentiation capability, transcriptomics and immunomodulatory potential of the iMSCs (iMSC012ARG, iMSC012TEX, iMSC013ARG and iMSC013TEX) and eMSCs (eMSC034ARG and eMSC034TEX) with nMSCs (nMSC012, nMSC013 and BM MSC). Based on the expression of all expressed genes, the data demonstrated that both iMSCs and eMSCs differentiated following TEX protocol are closer to nMSC with higher differentiation potentials but lower immunomodulatory properties than ARG.
Our data suggest that, following a careful selection and screening of donors, nMSCs from umbilical’s cord WJ can be reprogrammed into iPSCs providing an extensive source of material for differentiation into iMSCs of similar therapeutic potential as nMSC. iMSCs could be scaled up under GMP conditions and serve as an alternative to BM and MSCs from other sources for therapeutic purposes.

Date of Award	2019
Original language	English
Awarding Institution	King's College London
Supervisor	Dusko Ilic (Supervisor)