Skip to Content
Merck
  • Mesenchymal stromal cell proliferation, gene expression and protein production in human platelet-rich plasma-supplemented media.

Mesenchymal stromal cell proliferation, gene expression and protein production in human platelet-rich plasma-supplemented media.

PloS one (2014-08-15)
Paola Romina Amable, Marcus Vinicius Telles Teixeira, Rosana Bizon Vieira Carias, José Mauro Granjeiro, Radovan Borojevic
ABSTRACT

Platelet-rich plasma (PRP) is increasingly used as a cell culture supplement, in order to reduce the contact of human cells with animal-derived products during in vitro expansion. The effect of supplementation changes on cell growth and protein production is not fully characterized. Human mesenchymal stromal cells from bone marrow, adipose tissue and Wharton's Jelly were isolated and cultured in PRP-supplemented media. Proliferation, in vitro differentiation, expression of cell surface markers, mRNA expression of key genes and protein secretion were quantified. 10% PRP sustained five to tenfold increased cell proliferation as compared to 10% fetal bovine serum. Regarding cell differentiation, PRP reduced adipogenic differentiation and increased calcium deposits in bone marrow and adipose tissue-mesenchymal stromal cells. Wharton's Jelly derived mesenchymal stromal cells secreted higher concentrations of chemokines and growth factors than other mesenchymal stromal cells when cultured in PRP-supplemented media. Bone marrow derived mesenchymal stromal cells secreted higher concentrations of pro-inflammatory and pro-angiogenic proteins. Mesenchymal stromal cells isolated from adipose tissue secreted higher amounts of extracellular matrix components. Mesenchymal stromal cells purified from different tissues have distinct properties regarding differentiation, angiogenic, inflammatory and matrix remodeling potential when cultured in PRP supplemented media. These abilities should be further characterized in order to choose the best protocols for their therapeutic use.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
L-Ascorbic acid, FCC, FG
Supelco
L-Ascorbic acid, analytical standard
Sigma-Aldrich
L-Ascorbic acid, 99%
Sigma-Aldrich
L-Ascorbic acid, puriss. p.a., ACS reagent, reag. ISO, Ph. Eur., 99.7-100.5% (oxidimetric)
Sigma-Aldrich
L-Ascorbic acid, powder, suitable for cell culture, γ-irradiated
Sigma-Aldrich
L-Ascorbic acid, reagent grade
Sigma-Aldrich
Indomethacin, meets USP testing specifications
Sigma-Aldrich
L-Ascorbic acid, reagent grade, crystalline
Sigma-Aldrich
L-Ascorbic acid, BioXtra, ≥99.0%, crystalline
Sigma-Aldrich
Streptomycin sulfate salt, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Streptomycin sulfate salt, powder
Indomethacin, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Indomethacin, 98.5-100.5% (in accordance with EP)
Sigma-Aldrich
L-Ascorbic acid, ACS reagent, ≥99%
Sigma-Aldrich
L-Ascorbic acid, puriss. p.a., ≥99.0% (RT)
Supelco
Indomethacin, Pharmaceutical Secondary Standard; Certified Reference Material
Dexamethasone for peak identification, European Pharmacopoeia (EP) Reference Standard
Streptomycin sulfate, European Pharmacopoeia (EP) Reference Standard
Supelco
L-Ascorbic acid, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
USP
Indomethacin, United States Pharmacopeia (USP) Reference Standard
Supelco
Ciprofloxacin, VETRANAL®, analytical standard
Supelco
Ascorbic Acid, Pharmaceutical Secondary Standard; Certified Reference Material
USP
Ascorbic acid, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Ciprofloxacin, ≥98% (HPLC)
Sigma-Aldrich
Dexamethasone, powder, BioReagent, suitable for cell culture, ≥97%
Sigma-Aldrich
Alizarin Red S, certified by the BSC
Sigma-Aldrich
Transferrin human, powder, BioReagent, suitable for cell culture
Ascorbic acid, European Pharmacopoeia (EP) Reference Standard
Supelco
Ciprofloxacin, Pharmaceutical Secondary Standard; Certified Reference Material
Dexamethasone, European Pharmacopoeia (EP) Reference Standard