Trilineage Differentiation and Immunophenotyping of MSCs Isolated in Platelet Lysate as an FBS Alternative
Abstract
Mesenchymal stem cells (MSCs) are commonly used in regenerative medicine. Fetal bovine serum (FBS) is used to aid in the expansion of MSCs since it contains growth factors, but it also contains xenogens that are known to cause an inflammatory response. Platelet lysate (PL) has been suggested as an alternative to FBS because it contains growth factors and is a product of autologous or allogenic blood. However, the effectiveness of PL as an FBS substitute is not yet established. We hypothesized that equine MSCs grown in media containing 10% PL will maintain the same surface markers and trilineage differentiation capabilities as MSCs grown in media containing 10% FBS. MSCs were grown in three conditions: media containing FBS, media containing PL and heparin, and media containing FBS and heparin as a control. Trilineage differentiation was performed on each condition by supplementing MSC media with additives to induce differentiation into adipocytes, osteoblasts, and chondroblasts. Immunophenotyping was performed using flow cytometry for cell surface markers MHCII, CD90, and CD45. We observed lipid, calcium, and proteoglycan accumulation in all MSC lines. All but two cell lines were positive for CD90 (64.40% - 88.10%) and negative for CD45 (0.28% - 5.20%) and MHCII (0.985% - 1.280%). Based on our results, PL resulted in no changes in MSC characterization after expansion compared to FBS.
Subject
Mesenchymal stem cellsfetal bovine serum
platelet lysate
immunophenotyping
trilineage differentiation
Citation
Thornton, Rebecca Mayo; Hoathian, Michelle Brittany (2017). Trilineage Differentiation and Immunophenotyping of MSCs Isolated in Platelet Lysate as an FBS Alternative. Undergraduate Research Scholars Program. Available electronically from https : / /hdl .handle .net /1969 .1 /196694.