Platelet Lysate as Replacement for Fetal Bovine Serum in Mesenchymal Stromal Cell Cultures

Abstract

Mesenchymal stromal cells (MSC) emerged as highly attractive in cell-based regenerative medicine. Initially thought to provide cells capable of differentiation towards mesenchymal cell types (osteoblasts, chondrocytes, adipocytes etc.), by and by potent immunoregulatory and pro-regenerative activities have been discovered, broadening the field of potential applications from bone and cartilage regeneration to wound healing and treatment of autoimmune diseases. Due to the limited frequency in most tissue sources, ex vivo expansion of MSC is required compliant with good manufacturing practice (GMP) guidelines to yield clinically relevant cell doses. Though, still most manufacturing protocols use fetal bovine serum (FBS) as cell culture supplement to isolate and to expand MSC. However, the high lot-to-lot variability as well as risk of contamination and immunization call for xenogenic-free culture conditions. In terms of standardization, chemically defined media appear as the ultimate achievement. Since these media need to maintain all key cellular and therapy-relevant features of MSC, the development of chemically defined media is still - albeit highly investigated - only in its beginning. The current alternatives to FBS rely on human blood-derived components: plasma, serum, umbilical cord blood serum, and platelet derivatives like platelet lysate. Focusing on quality aspects, the latter will be addressed within this review.

Optimization of in vitro cultivation strategies for human adipocyte derived stem cells

“…A replacement with human serum derivatives has been reported previously and shown to support equal proliferation rates and multilineage differentiation capacity. 20,23 In our further studies we plan to approach those findings. …”

Standard Requirement of a Microbiological Quality Control Program for the Manufacture of Human Mesenchymal Stem Cells for Clinical Use

“…In the case of FBS, it is one of the most widely used reagents in the expansion of MSCs as cell culture supplement but the FBS presents a high variability between different batches as well as risk of contamination. As alternatives to this supplement, human factors from serum, plasma, or platelet derivatives have been suggested [36,37]. All of them must be included in the QCP, with stringent criteria of donor eligibility as well as sensitive viral NAT testing and serology, ensuring safety of the starting material for MSC manufacturing [37].…”

Fetal bovine serum (FBS): Past – present – future

“…In 2010, the revised EU Directive 2010/63/EU on the protection of animals used for scientific purposes (EU, 2010) was issued, also covering fetal forms of mammals. Furthermore, in recent years there have been increased efforts towards the establishment of human platelet lysates as alternatives to FBS for use as a cell culture supplement (Bieback, 2013;Burnouf et al, 2016;Gstraunthaler et al, 2015;Hemeda et al, 2014;Shih and Burnouf, 2015). In spring 2013, a severe case of fraudulent blending of FBS batches came to public attention (Gstraunthaler et al, , 2014.…”

“…Platelet (lysate) gels, the semisolid clot formed during platelet activation and fibrin formation, may become increasingly man serum-derived alternatives (Bieback, 2013;Burnouf et al, 2016;Gstraunthaler et al, 2015;Hemeda et al, 2014;Shih and Burnouf, 2015).…”