Human Platelet Lysate Can Replace Fetal Calf Serum as a Protein Source to Promote Expansion and Osteogenic Differentiation of Human Bone-Marrow-Derived Mesenchymal Stromal Cells

Abstract

Fetal calf serum (FCS) is frequently used as a growth factor and protein source in bone-marrow-derived mesenchymal stromal cell (BMSC) culture media, although it is a xenogenic product presenting multiple disadvantages including but not limited to ethical concerns. A promising alternative for FCS is human platelet lysate (hPL), which is produced out of human platelet concentrates and happens to be a stable and reliable protein source. In this study, we investigated the influence of hPL in an expansion medium (ESM) and an osteogenic differentiation medium (ODM) on the proliferation and osteogenic differentiation capacity of human BMSC. Therefore, we assessed population doublings during cell expansion, performed alizarin red staining to evaluate the calcium content in the extracellular matrix and determined the activity of alkaline phosphatase (ALP) as osteogenic differentiation correlates. The proliferation rate of BMSC cultured in ESM supplemented with hPL exceeded the proliferation rate of BMSC cultured in the presence of FCS. Furthermore, the calcium content and ALP activity was significantly higher in samples incubated in hPL-supplemented ODM, especially in the early phases of differentiation. Our results show that hPL can replace FCS as a protein supplier in cell culture media and does not negatively affect the osteogenic differentiation capacity of BMSC.

Protaetia brevitarsis seulensis Derived Protein Isolate with Enhanced Osteomodulatory and Antioxidative Property

“…Conversely, the expression of ALP and OPN decreased after 14 days of PPI treatment. The osteogenic marker genes are dynamically expressed during osteogenesis and often altered through protein supplementations [ 47 , 48 ]. Our results indicated that PPI was a potential modulator of the osteogenic gene expression, as compared to the well-established expression profile of osteogenic gene markers [ 49 ].…”

Use of conditioned media (CM) and xeno-free serum substitute on human adipose-derived stem cells (ADSCs) differentiation into urothelial-like cells

“…Several studies reported an increase in the differentiation potential of several types of MSCs towards the osteogenic, myofibroblastic, cardiomyogenic and adipogenic cell lineages in the presence of pHPL as a serum substitute ( Karadjian et al, 2020 ; Samuel et al, 2016 ; Chignon-Sicard et al, 2017 ; Homayouni Moghadam, Tayebi & Barzegar, 2016 ). On the other hand, pHPL decreased the ability of certain types of stem cells to differentiate into osteoblasts, chondrocytes and adipocytes ( Gruber et al, 2004 ; Lee et al, 2014 ; Chignon-Sicard et al, 2017 ).…”

Human Embryonic-Derived Mesenchymal Progenitor Cells (hES-MP Cells) are Fully Supported in Culture with Human Platelet Lysates

“…MSCs and hES-MP cells were grown in DMEM/F12+ Glutamax medium (Gibco, Grand Island, NY, USA), 1% penicillin/streptomycin (Gibco), and either 10% hPL (hPL–hES-MP and hPL–MSC treatments/cells) or 10% FBS (FBS–hES-MP and FBS–MSC treatments/cells). The decision to use 10% hPL was made after evaluating different hPL concentrations; our group typically uses 10% hPL in studies with bone marrow-derived MSCs, and this concentration has been investigated in other studies [ 30 , 31 , 32 ], which matches the typical concentration of FBS used for supplementation.…”

“…The cells were maintained at standard culture conditions, and the cell count was determined at the end of each passage using a Neubauer hemocytometer (Assistant, Munich, Germany). The number of PDs at each passage was then used to find the cumulative PDs (CPDs) using Equations (1) and (2), as previously described [ 28 , 32 ]: where N 0 and N H represent the number of cells seeded and the number of cells harvested (at the end of the expansion period), respectively, and n represents the number of passages.…”