Human platelet lysate as an alternative to fetal bovine serum for culture and endothelial differentiation of human amniotic fluid mesenchymal stem cells

Abstract

Human amniotic fluid (hAF) mesenchymal stem cells (MSCs) are commonly cultured in medium containing FBS. However, there are concerns about using animal serum in therapeutic applications due to the potential for immunogenic reactions and the risk of transmission of pathogens. For safety reasons, human platelet lysate (hPL) has been suggested as a replacement for FBS because it appears to be a natural source of growth factors. In this present study, it was investigated whether FBS could be substituted with hPL in hAF-MSCs culture without affecting their properties. Pooled hPL was generated by the freeze-thaw method. The concentration of hPL was selected after evaluation by MTT assay. The hAF-MSCs were cultured in FBS- or hPL-supplemented conditions and shared a fibroblast-like morphology. Cell proliferation assays showed that the growth characteristic of hAF-MSCs cultured in 10% hPL-supplemented media was similar to those cultured in 10% FBS-supplemented media. The expression of MSC markers did not differ between the cells cultured in the different conditions. The endothelial differentiation potential was also investigated. Reverse transcription-quantitative (RT-q)PCR revealed that induced cells supplemented with hPL showed an increase level of endothelial specific gene expression compared to the FBS-supplemented cells. Immunofluorescence analysis showed specific protein localization in both induced cell groups. Additionally, induced cells supplemented with hPL had the potential to form networks on Matrigel. This present study indicated that hPL could be used to culture and enhance the endothelial differentiation potential of hAF-MSCs.

Gelatin-Methacryloyl (GelMA) Formulated with Human Platelet Lysate Supports Mesenchymal Stem Cell Proliferation and Differentiation and Enhances the Hydrogel’s Mechanical Properties

“…Each GelMA solution was prepared in PBS with the addition of 0%, 2.5%, or 50% hPL (% v/v) or by adding hPL only (100% hPL), prior to the addition of the photoinitiator and the polymerization of the hydrogels. The specific hPL concentrations used in this study were selected because: (1) 2.5% hPL is the recommended concentration for 2D cell cultivation [33,34]; (2) 100% represents a complete substitution of PBS with hPL; (3) 50% hPL serves as an intermediate point between 2.5% and 100% hPL; and (4) 0% hPL functions as a control. Calcein-AM staining was performed to evaluate the influence of hPL addition on cell viability and spreading.…”

Platelet-derived extracellular vesicles promote osteoinduction of mesenchymal stromal cells

“…In this study, low PL doses have been used (2.6 μg of total protein/cm 2 , namely 0.02 ml of PL per 100 ml in the cell culture media in combination with a 1% EV-depleted FBS) while 10% of PL is normally used in substitution of FBS for MSC maintenance. 45 Further studies should be performed to evaluate the dose-effect response of PL and affirm the potential to use it as a FBS xeno-free substitute.…”

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

“…In particular Kandoi et al evidenced significantly faster cell growth of umbilical cord tissue-derived MSC in hPL-enriched cultures, reaching PDT of 20.95 h vs. PDT in FBS [ 5 ]. In contrast, Tancharoen et al revealed that amniotic fluid-derived MSC cultured in 10% FBS or hPL did not show differences in cell number and shared a fibroblast-like morphology [ 34 ]. In that regard, cell morphology has evolved as critical parameter predicting the expansion response to a particular cell supplement.…”