Human Platelet Lysate Supports Efficient Expansion and Stability of Wharton's Jelly Mesenchymal Stromal Cells via Active Uptake and Release of Soluble Regenerative Factors.

Bernardo Camacho,Carlos Medina,Mariana Cañas-Arboleda,Gustavo Salguero,Karl Beltrán

doi:10.3390/ijms21176284

Abstract

Manufacturing of mesenchymal stromal cell (MSC)-based therapies for regenerative medicine requires the use of suitable supply of growth factors that enhance proliferation, cell stability and potency during cell expansion. Human blood derivatives such as human platelet lysate (hPL) have emerged as a feasible alternative for cell growth supplement. Nevertheless, composition and functional characterization of hPL in the context of cell manufacturing is still under investigation, particularly regarding the content and function of pro-survival and pro-regenerative factors. We performed comparative analyses of hPL, human serum (hS) and fetal bovine serum (FBS) stability and potency to support Wharton’s jelly (WJ) MSC production. We demonstrated that hPL displayed low inter-batch variation and unique secretome profile that was not present in hS and FBS. Importantly, hPL-derived factors including PDGF family, EGF, TGF-alpha, angiogenin and RANTES were actively taken up by WJ-MSC to support efficient expansion. Moreover, hPL but not hS or FBS induced secretion of osteoprotegerin, HGF, IL-6 and GRO-alpha by WJ-MSC during the expansion phase. Thus, hPL is a suitable source of factors supporting viability, stability and potency of WJ-MSC and therefore constitutes an essential raw material that in combination with WJ-MSC introduces a great opportunity for the generation of potent regenerative medicine products.

Highlights

Application of cell therapies based on the use of mesenchymal stromal cells (MSC) has gained a major place among current approaches for regenerative medicine worldwide
Setting up an optimized workflow for the generation of reproducible starting material required for cell production, is critical to ensure consistent and reliable growth rates of Wharton’s jelly (WJ)-MSC for clinical scaling
An essential quality standard required for the use of human platelet lysate (hPL) as culture supplement for expansion of good manufacturing practices (GMP)-grade WJ-MSC is linked to its stability under different storage conditions

Summary

Introduction

Application of cell therapies based on the use of mesenchymal stromal cells (MSC) has gained a major place among current approaches for regenerative medicine worldwide. Advances in the adoption of new technologies and procedures oriented to the manufacturing of MSC-based medicinal products have been pivotal to speed-up translation into clinical settings of a wide variety of therapeutic tools from tissue engineering to immunotherapy In this context, optimizing cell manufacturing workflow is a critical step to obtain better and more effective therapies by ensuring high yield, viability, reproducibility and safety of the active cellular product. Wharton’s jelly (WJ) MSC are being intensively explored as “off-the-shelf” cell therapy for a wide array of chronic degenerative disorders including autoimmune and orthopedic conditions In this context, generation of fully characterized, already-available WJ-MSC allogeneic cell banks has broadened the application potential of MSC products, thereby accelerating their clinical translation for regenerative medicine [14,15]. We hypothesized that hPL provides critical factors that are taken up by WJ-MSC to enhance their regenerative potential via constitutive secretion of pro-survival and immunomodulatory signals

Production of hPL and hS for Culture of WJ-MSC

Cytokine and Growth Factor Stability Present in hPL

Materials and Methods

Isolation and Culture of WJ-MSC

WJ-MSC Immunophenotype Analyses

Assessment T Lymphocyte Suppression by WJ-MSC

Determination of Cytokine and Growth Factor Levels by Multiplex Bead Analysis

Statistical Analysis