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KMID : 1100720140340010043
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Annals of Laboratory Medicine
2014 Volume.34 No. 1 p.43 ~ p.50
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Effects of Platelet Lysate Preparations on the Proliferation of HaCaT Cells
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Baik Sae-Yun
Lim Young-Ae
Kang Seon-Joo
Ahn Sun-Hyun
Lee Wee-Gyo
Kim Chul-Ho
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Abstract
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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 1×1012/L and 2×1012/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 2×1012/L than 1×1012/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 1×1012/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.
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KEYWORD
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Platelet lysate, Cell culture, Freeze-thaw, Growth factor, Cell proliferation
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