Abstract
Human embryonic stem cell-derived mesenchymal progenitor (hES-MP) cells are mesenchymal-like cells, derived from human embryonic stem cells without the aid of feeder cells. They have been suggested as a potential alternative to mesenchymal stromal cells (MSCs) in regenerative medicine due to their mesenchymal-like proliferation and differentiation characteristics. Cells and cell products intended for regenerative medicine in humans should be derived, expanded and differentiated using conditions free of animal-derived products to minimize risk of animal-transmitted disease and immune reactions to foreign proteins. Human platelets are rich in growth factors needed for cell culture and have been used successfully as an animal serum replacement for MSC expansion and differentiation. In this study, we compared the proliferation of hES-MP cells and MSCs; the hES-MP cell growth was sustained for longer than that of MSCs. Growth factors, gene expression, and surface marker expression in hES-MP cells cultured with either human platelet lysate (hPL) or fetal bovine serum (FBS) supplementation were compared, along with differentiation to osteogenic and chondrogenic lineages. Despite some differences between hES-MP cells grown in hPL- and FBS-supplemented media, hPL was found to be a suitable replacement for FBS. In this paper, we demonstrate for the first time that hES-MP cells can be grown using platelet lysates from expired platelet concentrates (hPL).
Highlights
The field of regenerative medicine has rapidly expanded in recent years
The SOX9 expression was observed to be higher in human platelet lysate (hPL) chondrocytes at earlier time points, but at day 28, a 4.27 ± 0.04 fold greater increase was observed in the SOX9 expression in the fetal bovine serum (FBS) chondrocytes than in the hPL chondrocytes, after which the expression remained higher in FBS chondrocytes and 3.97 ± 0.08 fold on day 35; p ≤ 0.001; n = 3)
Discussion Human embryonic stem cell-derived mesenchymal progenitor (hES-MP) cells have been reported to have similar characteristics and bioactivity to mesenchymal stromal cells (MSCs) [22]. They have been suggested as a potential off-the-shelf product that can be handled and used in an identical manner to MSCs [22,34]. hPL has been widely described as a promising culture supplement for MSCs, but the evidence regarding hES-MP cells has been lacking
Summary
The field of regenerative medicine has rapidly expanded in recent years. Various types of somatic stem cells, as well as embryonic and induced pluripotent stem cells, have been evaluated for their regenerative abilities [1,2,3,4].Adult human mesenchymal stromal cells (MSCs) have been studied extensively and are considered promising candidates for regenerative therapies due to their differentiation potential and immunomodulatory function [5,6,7]. Several limitations are associated with MSCs that may hamper their use in regenerative medicine. These include inter-donor variability, variable isolation protocols, which commonly result in a heterogeneous population of cells, reduced proliferation capacity with cell age, and impairment of biological characteristics following long-term in vitro expansion [5,8,9,10,11]. Various protocols are available that describe the derivation of cells with high MSC resemblance from ESCs [12,13,14,15,16,17,18], with the aim of combining the regenerative potential of MSCs with the long-term proliferation of ESCs. ESCs are commonly grown on feeder cells, such as mouse fibroblast cells [19]. Animal-free conditions are, recommended when deriving and growing cells intended for human use [1,20,21]
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