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.

Maria Karadjian,Christopher Essers,Raban Heller,Anne-Sophie Senger,Fabian Westhauser,Joerg Fellenberg,Sebastian Wilkesmann,Rolf Simon

doi:10.3390/cells9040918

Maria Karadjian, Christopher Essers + Show 6 more

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https://doi.org/10.3390/cells9040918

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Journal: Cells	Publication Date: Apr 9, 2020
Citations: 17	License type: CC BY 4.0

Affiliation: University Hospital Heidelberg, Heidelberg University

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.

Highlights

Despite some limitations, fetal calf serum (FCS) is still the most common source of proteins and growth factors in cell culture media [1]
As most available studies on human platelet lysate (hPL) assess its effects on mesenchymal stromal cells (MSC) during the expansion period, we focused on the effects of hPL as part of osteogenic differentiation medium (ODM) on the kinetics of bone-marrow-derived mesenchymal stromal cell (BMSC) osteogenic differentiation by evaluating alkaline phosphatase (ALP) activity and calcium deposition as osteogenic differentiation markers at multiple time points
Based on the assays performed, the results presented in this study confirm the results of previous studies that can be explained by the higher amount of growth factors in hPL, favoring cell proliferation and leading to a higher absolute growth factor amount in cell culture media when supplemented in the same concentration as Fetal calf serum (FCS) [11]

Summary

Introduction

Fetal calf serum (FCS) is still the most common source of proteins and growth factors in cell culture media [1]. The conditions of FCS production regarding animal welfare have to be viewed critically, especially since production circumstances often remain unclear and information gathering is reported to be complicated [12]. This raises the question whether an alternative protein source standard in cell culture media is required.

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