Predicting and Promoting Human Bone Marrow MSC Chondrogenesis by Way of TGFβ Receptor Profiles: Toward Personalized Medicine.

René Rothweiler,David Kubosch,Fabian Duttenhoefer,Brian Johnstone,Martin James Stoddart,Valentina Basoli,Rainer Schmelzeisen,Mauro Alini

doi:10.3389/fbioe.2020.00618

Abstract

The use of human mesenchymal stromal cells (hMSCs) for cartilage regeneration has been hampered by the inherent donor variation of primary monolayer expanded cells. Although CD markers are typically used to characterize cell populations, there is no correlation between CD marker profile and functional outcomes. Therefore, we aimed to discover novel predictive MSC chondrogenesis markers. The chondrogenic potential of primary human bone marrow MSCs (hBMSCs) over multiple passages was assessed by standard pellet culture. We confirmed that the ratio of TGFβ-RI/TGFβ-RII at the time of cell recovery from the tissue culture plastic reliably predicted chondrogenic potential. Furthermore, it is possible to prospectively characterize any human BMSC cell population as responders or non-responders with respect to chondrogenic differentiation potential. Transient increase of the ratio with siRNA knockdown of TGFβ-RII reproducibly recovered the chondrogenic differentiation ability of non-responsive MSCs. Together this offers an opportunity to produce a more functionally characterized cell population for use in autologous cartilage repair therapies.

Highlights

Despite the promise of human bone marrow derived stromal cells in the field of regenerative medicine, assays predictive of cell function have remained elusive
We compared the means and standard deviations up to passage 7 and investigated whether changes in the expression profiles of TGFβRs and BMP-Rs during cell passaging could be associated with changing chondrogenic potential (Figure 1)
There was a high standard deviation (SD) among the donors, and several receptors displayed an increase in SD with increasing passaging, leading us to investigate each receptor at the individual donor level

Summary

Introduction

Despite the promise of human bone marrow derived stromal cells (hBMSCs) in the field of regenerative medicine, assays predictive of cell function have remained elusive. The consequence of our current incomplete picture of the causes of MSC variability is that to date, it is not possible to predict the outcome of chondrogenic differentiation of a specific donor. This is a clinical challenge as cell based therapies are expensive. Most marker profiles are similar for all cells of mesenchymal origin (Whitney et al, 2009) but hMSCs from different origins have been shown to retain epigenetic memory and display functional differences in vivo. For this reason new methods for predicting the functional potential of hMSCs are urgently needed (McLeod and Mauck, 2017)

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