Effects of platelet lysate preparations on the proliferation of HaCaT cells.

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 1x10(12)/L and 2x10(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 2x10(12)/L than 1x10(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 1x10(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.