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TRANSPLANTATION AND CELLULAR ENGINEERING: Adipose tissue mesenchymal stem cell expansion in animal serum‐free medium supplemented with autologous human platelet lysate
Author(s) -
Blande Ivan Souza,
Bassaneze Vinicius,
LaviniRamos Carolina,
Fae Kellen Cristhina,
Kalil Jorge,
Miyakawa Ayumi Aurea,
Schettert Isolmar Tadeu,
Krieger Jose Eduardo
Publication year - 2009
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/j.1537-2995.2009.02346.x
Subject(s) - platelet lysate , mesenchymal stem cell , fetal bovine serum , stem cell , adipose tissue , andrology , immunology , tissue engineering , transplantation , platelet rich plasma , cell therapy , biology , microbiology and biotechnology , platelet , chemistry , in vitro , medicine , biomedical engineering , surgery , endocrinology , biochemistry
BACKGROUND: Mesenchymal stem cells (MSCs) have been considered for human regenerative therapy applications, and safe culture and expansion protocols are needed especially in the context of interspecies contamination. Human platelet lysate (PL) has been proposed as animal serum substitute during in vitro MSC expansion. In this work, a simplified and efficient method to obtain autologous PL to replace animal serum in cell culture applications is described. STUDY DESIGN AND METHODS: PL obtained by freezing and centrifugation procedures was tested as medium supplement for human adipose mesenchymal stem cell (hASC) culture. Differential proliferation, immunophenotypic changes, and differentiation under PL or fetal bovine serum (FBS) were assessed. RESULTS: In contrast to 10% FBS supplementation, cell population doubling time was significantly lower when hASCs were cultured with the same concentration of PL (PL 22.9 ± 1.5 hr vs. FBS 106.7 ± 6.5 hr, t test, p < 0.05). Furthermore, hASCs maintained with 2.5% PL supplementation also showed satisfactory results. Immunophenotypic analysis revealed no differences between hASCs cultivated with PL or FBS supplementation and both cultures retained the potential to differentiate into adipose cells. These results demonstrate that autologous PL obtained from the same donor can be used as animal serum substitute in hASC culture. CONCLUSIONS: Taken together, evidence is provided that platelets provided by a single donor are sufficient to obtain PL for hASC propagation for clinical‐scale applications mitigating the potential untoward side effects associated with the use of animal‐derived reagents.