Effects of Platelet Lysate Preparations on the Proliferation of HaCaT Cells
Annals of Laboratory Medicine
;
: 43-50, 2014.
Article
in English
| WPRIM
| ID: wpr-193129
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Blood Platelets
/
Platelet-Derived Growth Factor
/
Cell Line
/
Culture Media
/
Vascular Endothelial Growth Factor A
/
Cell Proliferation
/
Epidermal Growth Factor
Type of study:
Practice guideline
Limits:
Humans
Language:
English
Journal:
Annals of Laboratory Medicine
Year:
2014
Type:
Article
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