The production method affects the efficacy of platelet derivatives to expand mesenchymal stromal cells in vitro

Journal of Translational Medicine

Research

Eliana Amati¹  Sabrina Sella¹  Giuseppe Astori¹  Marco Ruggeri¹  Katia Chieregato²  Martina Bernardi²  Mario Rassu³  Mario Mazzucato⁴  Francesco Agostini⁴  Cristina Durante⁴  Elisabetta Lombardi⁴ 

[1] Advanced Cellular Therapy Laboratory, Hematology Unit, Vicenza Hospital, Vicenza, Italy;Advanced Cellular Therapy Laboratory, Hematology Unit, Vicenza Hospital, Vicenza, Italy;Hematology Project Foundation, Contrà S. Francesco 41, Vicenza, Italy;Department of Microbiology, San Bortolo Hospital, Via Rodolfi 37, 36100, Vicenza, Italy;Stem Cell Collection and Processing Unit, CRO National Cancer Institute-IRCCS Aviano, Aviano, Italy;

关键词: Platelet lysate;    Platelet releasate;    Mesenchymal stromal cells;    Mesenchymal stem cells;    Ex vivo expansion;    Fetal bovine serum;   

DOI  :  10.1186/s12967-017-1185-9

 received in 2016-09-16, accepted in 2017-04-14,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundThe use of fetal bovine serum (FBS) as a media supplement for the ex vivo expansion of bone-marrow derived mesenchymal stromal cells (BM-MSC) has been discouraged by regulatory agencies, due to the risk of transmitting zoonoses and to elicit immune reactions in the host once transplanted. Platelet derivatives are valid FBS substitutes due to their content of growth factors that can be released disrupting the platelets by physical methods or physiological stimuli. We compared platelet derivatives produced by freezing/thawing (platelet lysates, PL) or after CaCl2 activation (platelet releasate surnatant rich in growth factors, PR-SRGF) for their content in growth factors and their ability to support the ex vivo expansion of BM-MSC.MethodsThe cytokine content in the two platelet derivatives was evaluated. BM-MSC were expanded in complete medium containing 10, 7.5 and 5% PL or PR-SRGF and the cell phenotype, clonogenic capacity, immunomodulation properties and tri-lineage differentiation potential of the expanded cells in both media were investigated.ResultsThe concentration of PDGF-AB, PDGF-AA, PDGF-BB in PR-SRGF resulted to be respectively 5.7×, 1.7× and 2.3× higher compared to PL. PR-SRGF promoted a higher BM-MSC proliferation rate compared to PL not altering BM-MSC phenotype. Colony forming efficiency of BM-MSC expanded in PR-SRGF showed a frequency of colonies significantly higher than cells expanded in PL. BM-MSC expanded in PL or PR-SRGF maintained their immunomodulatory properties against activated lymphocytes even if BM-MSC expanded in FBS performed significantly better.ConclusionsThe method used to release platelet factors significantly affects the enrichment in growth factors and overall product performance. The standardization of the production process of platelet derivatives and the definition of their release criteria requires further investigation.

【 授权许可】

CC BY   
© The Author(s) 2017