Effect of Human Platelet Lysate in Differentiation of Wharton’s Jelly Derived Mesenchymal Stem Cells

Abstract

Background: Mesenchymal stem cells (MSCs) are highly preferred in clinical therapy for repair and regeneration of diseased tissues for their multipotent properties. Conventionally, MSCs have been cultured in media supplemented with animal derived serum, however, it is ideal to expand MSCs in media containing supplements of human origin for clinical therapy. Currently, a number of human derived products are being studied as an alternative to animal sources. Amongst these, platelet lysate (PL) has gained interest in the culture of MSCs without affecting their phenotypic property. Objective: In this study, we used various concentration of PL (2.5, 5, 7.5 & 10%) in the growth medium of MSCs to identify the least concentration of PL that could be an effective alternative to animal products. Methods: MSCs were isolated from Wharton’s Jelly by using explant method and expanded in various concentration of PL supplemented medium against the standard FBS containing medium. WJ-MSCs were characterised as per the minimal criteria proposed by International Society for Cell therapy (ISCT), Proliferation study by BrdU assay, gene expression study by qRT-PCR, sterility test for bacteria, Mycoplasma by PCR and endotoxin detection by LAL assay. Results: Whartons jelly derived MSCs (WJ-MSCs) cultured using standard medium supplemented with various concentration of PL exhibited enhanced proliferation and differentiation potential, unaltered immunophenotypic property and genetic stability when compared with the commercial medium containing 10% FBS. Conclusion: The least concentration of PL for an ideal expansion of MSCs was found to be 2.5% and was comparable to FBS.

Effect of Human Platelet Lysate as Cultivation Nutrient Supplement on Human Natal Dental Pulp Stem Cell In Vitro Expansion

“…Interestingly, hPL showed high expression of all tested markers (<90%). These results differ from other studies where no effect of the medium supplement was observed [ 44 , 45 , 46 ]. There was no statistical difference in mesenchymal stem cell markers (CD29, CD44, CD73, CD90) and stromal-associated markers (CD13 and CD166).…”

“…hPL-cultivated hNDP-SCs showed approximately two times shorter PDT (22.65 ± 0.10 h) compared with the FBS-cultivated group (44.69 ± 1.36 h), while the PDs per passage were the same (3.97 ± 0.35 vs. 3.98 ± 0.38). Our results are comparable with other studies [ 26 , 36 , 39 , 40 , 43 , 44 ]. At the end of cell growth, we observed the prolongation of population doubling time in the hPL-treated group.…”

Expanding applications of allogeneic platelets, platelet lysates, and platelet extracellular vesicles in cell therapy, regenerative medicine, and targeted drug delivery

“…It was observed in some studies that supplementation with HPLs versus FBS favors MSC differentiation into chondrocytes [ 126 , 128 , 130 , 169 ]. HPLs are also a suitable supplement to expand MSCs from other tissue sources such as Wharton jelly [ 126 , 170 ], umbilical cords [ 171 – 174 ], amniotic fluid [ 175 , 176 ], dental pulp [ 177 – 179 ], periodontal ligaments [ 180 ], and others [ 121 ]. HPLs are therefore emerging as a universal supplement for in vitro xeno-free expansion of MSCs [ 12 , 121 , 181 ] for clinical transplantation.…”

An optimized procedure for preparation of conditioned medium from Wharton’s jelly mesenchymal stromal cells isolated from umbilical cord

“…Media supplemented with FBS are widely used to provide a supportive environment for isolation and expansion of MSCs from various sources, despite notorious practical, clinical, and ethical concerns over FBS use, essentially due to the presence of undesirable toxins, pathogen agents and/or xenogeneic proteins ( Shahdadfar et al, 2005 ; van der Valk and Gstraunthaler, 2017 ). For this reason, in order to develop a clinically suitable and hazardous risk-free protocol for the isolation of Wharton’s jelly-derived MSCs (WJ-MSCs) from umbilical cords, many reports showed the potential of human platelet lysate (hPL) in enhancing MSCs recovery and propagation ( de Soure et al, 2017 ; Becherucci et al, 2018 ; Kandoi et al, 2018 ; Vennila et al, 2019 ; Liao et al, 2021 ).…”