Comparing the Effect of TGF-β Receptor Inhibition on Human Perivascular Mesenchymal Stromal Cells Derived from Endometrium, Bone Marrow and Adipose Tissues.

Shanti Gurung,Anna Rosamilia,Jerome A Werkmeister,Marian Sturm,Caroline E Gargett,Daniela Ulrich

doi:10.3390/jpm10040261

Abstract

Rare perivascular mesenchymal stromal cells (MSCs) with therapeutic properties have been identified in many tissues. Their rarity necessitates extensive in vitro expansion, resulting in spontaneous differentiation, cellular senescence and apoptosis, producing therapeutic products with variable quality and decreased potency. We previously demonstrated that A83-01, a transforming growth factor beta (TGF-β) receptor inhibitor, maintained clonogenicity and promoted the potency of culture-expanded premenopausal endometrial MSCs using functional assays and whole-transcriptome sequencing. Here, we compared the effects of A83-01 on MSCs derived from postmenopausal endometrium, menstrual blood, placenta decidua-basalis, bone marrow and adipose tissue. Sushi-domain-containing-2 (SUSD2+) and CD34+CD31−CD45− MSCs were isolated. Expanded MSCs were cultured with or without A83-01 for 7 days and assessed for MSC properties. SUSD2 identified perivascular cells in the placental decidua-basalis, and their maternal origin was validated. A83-01 promoted MSC proliferation from all sources except bone marrow and only increased SUSD2 expression and prevented apoptosis in MSCs from endometrial-derived tissues. A83-01 only improved the cloning efficiency of postmenopausal endometrial MSCs (eMSCs), and expanded adipose tissue MSCs (adMSCs) underwent significant senescence, which was mitigated by A83-01. MSCs derived from bone marrow (bmMSCs) were highly apoptotic, but A83-01 was without effect. A83-01 maintained the function and phenotype in MSCs cultured from endometrial, but not other, tissues. Our results also demonstrated that cellular SUSD2 expression directly correlates with the functional phenotype.

Highlights

Perivascular mesenchymal stromal cells (MSCs) are a rare population of cells that self-renew and maintain homeostasis of the tissues in which they reside [1,2]
This study demonstrated that A83-01 significantly promoted MSC proliferation, increased the percentage of SUSD2+ MSCs and decreased apoptotic cells, especially in cultures derived from endometrium-derived tissues—postmenopausal endometrium, placental decidua basalis and menstrual blood
We demonstrated that A83-01, a TGF-β receptor inhibitor, increased the proliferation of long-term cultured MSCs from the postmenopausal endometrium, menstrual blood, placental decidua basalis and adipose tissue in a similar manner to premenopausal endometrial MSCs (eMSCs) [46], indicating that the endogenous synthesis of TGF-β had a negative effect on MSC growth during culture expansion in serum-free medium (SFM)

Summary

Introduction

Perivascular mesenchymal stromal cells (MSCs) are a rare population of cells that self-renew and maintain homeostasis of the tissues in which they reside [1,2]. They are multipotent [3] and have immunomodulatory properties mediated by a paracrine mechanism through their secretory profile [4]. There is recent concern about the possibility of an acute antidonor immune response against infused MSCs following repeated allo-MSC administration [7,8,9,10]. The exciting discovery of MSCs in most tissues of the body has reinvigorated the discussion on using MSCs as an autologous cell source for therapies [13]

Objectives

Methods

Results

Discussion

Conclusion