Adipose tissue mesenchymal stem cell expansion in animal serum-free medium supplemented with autologous human platelet lysate

Autor(es)

Blande, Ivan Souza ; Bassaneze, Vinicius ; Lavini-Ramos, Carolina ; Fae, Kellen Cristhina ; Kalil, Jorge ; Miyakawa, Ayumi Aurea ; Schettert, Isolmar Tadeu ; Krieger, Jose Eduardo ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Sch Med, Heart Inst InCor, Lab Genet & Mol Cardiol LIM 13, BR-05403000 Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Transfusion; v. 49, n. 12, p. 2680-2685, DEC 2009.
• Citações Web of Science: 64

Resumo

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. (AU)

• Processo FAPESP: 08/52436-9 - Reprogramação dirigida de células mesenquimais de tecido adiposo em células-tronco pluripotentes e cardiomiócitos
• Beneficiário: José Eduardo Krieger
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 05/57591-4 - Caracterizacao da influencia do shear stress na diferenciacao de celulas mesenquimais de tecido adiposo em celulas endoteliais.
• Beneficiário: Vinícius Bassaneze
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 06/55707-8 - Substituicao do soro fetal bovino (fbs) por lisado de plaquetas (pl) no meio de cultura: estudo da proliferacao e permanencia do estado de quiescencia de celulas mesenquimais do tecido adiposo...
• Beneficiário: Ivan Souza Blande
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica