Effects of platelet lysate preparations on the proliferation of HaCaT cells.
Ann Lab Med
; 34(1): 43-50, 2014 Jan.
Article
em En
| MEDLINE
| ID: mdl-24422195
ABSTRACT
BACKGROUND:
Standard protocols are lacking for the preparation of platelet lysates (PL) as an alternative to using fetal bovine serum as a cell culture supplement. This study aimed to establish optimum conditions for preparing PL for use in cell cultures.METHODS:
Cell density in three pooled platelet concentrates (PC) were adjusted to 1×10(12)/L and 2×10(12)/L. PL was prepared from PC by 1 to 3 freeze-thaw (FT) cycles. HaCaT cells were cultured in media supplemented with 5% or 10% PL. Cell numbers were estimated using a Cell Counting Kit-8 (CCK-8; Dojindo Laboratories, Japan). Growth factors were quantified by using the Luminex 200 system (Luminex Corporation, USA).RESULTS:
Cell proliferation rates in the presence of PLs were similar when prepared from PCs of both cell densities. The rates were higher in media containing 5% PL than 10% PL when prepared by two FT cycles. Concentrations of vascular endothelial growth factor (VEGF), platelet-derived growth factor-AB/BB (PDGF-AB/BB), PDGF-AA, and epidermal growth factor (EGF) were significantly higher in PL prepared from PC with a cell density of 2×10(12)/L than 1×10(12)/L PC. However, only VEGF and PDGF-AA concentrations in PLs were correlated with HaCaT cell counts.CONCLUSIONS:
The 5% PL from PC with a cell density of 1×10(12)/L prepared by two FT cycles treatment was the most effective condition that supported steady HaCaT cell proliferation. Our finding may be useful for preparing PL-supplemented cell culture media.Palavras-chave
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Base de dados:
MEDLINE
Assunto principal:
Plaquetas
Tipo de estudo:
Guideline
Limite:
Humans
Idioma:
En
Ano de publicação:
2014
Tipo de documento:
Article