Growth factors-based beneficial effects of platelet lysate on umbilical cord-derived stem cells and their synergistic use in osteoarthritis treatment

Li Yan,Bo Yan,Thomas Efferth,Wenxi Du,Li Zhou,Peijian Tong,Fucun Liu,Letian Shan,Qiang Yuan,Lei Zhang

doi:10.1038/s41419-020-03045-0

Abstract

Poor viability of mesenchymal stem cells (MSCs) at the transplanted site often hinders the efficacy of MSCs-based therapy. Platelet lysate (PL) contains rich amounts of growth factors, which benefits cell growth. This study aimed to explore how human PL benefits umbilical cord-derived MSCs (huc-MSCs), and whether they have synergistic potential in osteoarthritis (OA) treatment. As quality control, flow cytometry and specific staining were performed to identify huc-MSCs, and ELISA was used to quantify growth factors in PL. CCK-8 and flow cytometry assays were performed to evaluate the effects of PL on the cell viability and cell cycle progression of huc-MSCs. Wound healing and transwell assays were conducted to assess the migration of huc-MSCs. RNA sequencing, real time PCR, and Western blot assays were conducted to explore the growth factors-based mechanism of PL. The in vitro results showed that PL significantly promoted the proliferation, cell cycle, and migration of huc-MSCs by upregulating relevant genes/proteins and activating beclin1-dependent autophagy via the AMPK/mTOR signaling pathway. The main growth factors (PDGF-AA, IGF-1, TGF-β, EGF, and FGF) contributed to the effects of PL in varying degrees. The in vivo data showed that combined PL and huc-MSCs exerted significant synergistic effect against OA. The overall study determined the beneficial effects and mechanism of PL on huc-MSCs and indicated PL as an adjuvant for huc-MSCs in treating OA. This is the first report on the growth factors-based mechanism of PL on huc-MSCs and their synergistic application. It provides novel knowledge of PLʹs roles and offers a promising strategy for stem cell-based OA therapy by combining PL and huc-MSCs.

Highlights

Mesenchymal stem cells (MSCs) become an attractive and promising therapeutic tool for cell therapy and regenerative medicine, due to their self-renewal capacity, multilineage differentiation potential, paracrine effects, and immunomodulatory properties, which can promote all regenerative stages of damaged tissues
Huc-MSCs identification As shown in Fig. 1A, the umbilical cord-isolated cells showed fibroblast-like morphology with plastic adherent properties and expressed the surface markers CD73 (>99%), CD90 (>99%), and CD105 (>98%), but not CD14 (
The main innovation of this paper is the finding of Platelet lysate (PL)′s beneficial effects on huc-MSCs and its mechanism of growth factors-based action, which differs from previous studies focusing mainly on the potential of PL as substitute for fetal bovine serum (FBS) in MSCs expansion

Summary

Introduction

Mesenchymal stem cells (MSCs) become an attractive and promising therapeutic tool for cell therapy and regenerative medicine, due to their self-renewal capacity, multilineage differentiation potential, paracrine effects, and immunomodulatory properties, which can promote all regenerative stages of damaged tissues. Human umbilical cordderived MSCs (huc-MSCs) are noncontroversial compared to embryonic stem cells and induced pluripotent stem cells[2,4]. Huc-MSCs possess higher proliferative potential than MSCs from other sources[5]. Yan et al Cell Death and Disease (2020)11:857 of teratomas or tumors, and strong immunomodulatory capacities[6]. The FDA has registered dozens of clinical trials on the transplantation of huc-MSCs for treating refractory diseases, such as knee osteoarthritis (OA)[2,8]

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