title: 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 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.
authors.affiliation: Green Cross Laboratories, Yongin, Korea.
authors.forename: Sae Yun
authors.initials: SY
authors.lastname: Baik
authors.affiliation: Department of Laboratory Medicine, School of Medicine, Ajou University, Suwon, Korea.
authors.forename: Young Ae
authors.initials: YA
authors.lastname: Lim
authors.affiliation: Department of Laboratory Medicine, School of Medicine, Ajou University, Suwon, Korea.
authors.forename: Seon Joo
authors.initials: SJ
authors.lastname: Kang
authors.affiliation: Department of Laboratory Medicine, School of Medicine, Ajou University, Suwon, Korea.
authors.forename: Sun Hyun
authors.initials: SH
authors.lastname: Ahn
authors.affiliation: Department of Laboratory Medicine, School of Medicine, Ajou University, Suwon, Korea.
authors.forename: Wee Gyo
authors.initials: WG
authors.lastname: Lee
authors.affiliation: Department of Otolaryngology, School of Medicine, Ajou University, Suwon, Korea.
authors.forename: Chul Ho
authors.initials: CH
authors.lastname: Kim
mesh_terms: D001792:Blood Platelets / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D003470:Culture Media / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D010982:Platelet-Derived Growth Factor / Q000737:chemistry / Q000494:pharmacology; D042461:Vascular Endothelial Growth Factor A / Q000737:chemistry / Q000494:pharmacology
chemical_list: D003470:Culture Media; D010982:Platelet-Derived Growth Factor; D042461:Vascular Endothelial Growth Factor A; C075278:platelet-derived growth factor A; C095477:platelet-derived growth factor AB; D004815:Epidermal Growth Factor
keywords: Cell culture; Cell proliferation; Freeze-thaw; Growth factor; Platelet lysate