Enhanced ex vivo expansion of human adipose tissue-derived mesenchymal stromal cells by fibroblast growth factor-2 and dexamethasone.
In the present study, we investigated the ex vivo expansion of human adipose tissue-derived mesenchymal stromal cells (ATSCs) to identify factors that promoted efficient expansion while preserving stem cell potential. We examined several growth factors and steroids, and found that the combination of a low concentration of fibroblast growth factor-2 (FGF-2) (1 ng/mL) and dexamethasone (DEX) or betamethasone (BET) enhanced the proliferation of ATSCs by approximately 30-60% as compared to control. Enhanced proliferation under these conditions was confirmed using ATSCs isolated from three independent donors. ATSCs that were expanded in the presence of FGF-2 and DEX for 5 days were capable of differentiating into either osteoblastic or adipogenic cells, and the cells were positive for the mesenchymal stem cell markers such as CD29, CD44, CD90, CD105, and CD146, suggesting that the stem cell potential of the ATSCs was preserved. Analysis of signaling pathway revealed that tyrosine phosphorylation of Src kinase was dramatically increased in response to FGF-2 and DEX, suggesting the involvement of Src-dependent pathways in the stimulatory mechanism of proliferation of ATSCs by FGF-2 and DEX. Moreover, Src family kinase inhibitors (SU6656 and Src kinase inhibitor I) substantially reduced the FGF-2 and DEX-induced proliferation of ATSCs. SU6656 also inhibited the osteogenic and adipogenic differentiation of ATSCs. The results of the current study demonstrate that FGF-2 in combination with DEX stimulates the proliferation and osteoblastic and adipogenic differentiation of ATSCs through a Src-dependent mechanism, and that FGF-2 and DEX promote the efficient ex vivo expansion of ATSCs.
Lee
Sun-Young
SY
Skeletal Diseases Genome Research Center, Kyungpook National University Hospital, Daegu, Republic of Korea.
Lim
Jiwon
J
Khang
Gilson
G
Son
Youngsook
Y
Choung
Phil-Hoon
PH
Kang
Shin-Sung
SS
Chun
So Young
SY
Shin
Hong-In
HI
Kim
Shin-Yoon
SY
Park
Eui Kyun
EK
D050156:Adipogenesis / Q000187:drug effects; D000273:Adipose Tissue / Q000166:cytology*; D015415:Biomarkers / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003907:Dexamethasone / Q000494:pharmacology*; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000201:enzymology; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects; D019000:Phosphotyrosine / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D017154:Stromal Cells / Q000166:cytology* / Q000187:drug effects* / Q000201:enzymology; D013449:Sulfonamides / Q000494:pharmacology; D014019:Tissue Donors; D019061:src-Family Kinases / Q000037:antagonists & inhibitors
D015415:Biomarkers; D007211:Indoles; D047428:Protein Kinase Inhibitors; C416927:SU 6656; D013449:Sulfonamides; D016222:Fibroblast Growth Factor 2; D019000:Phosphotyrosine; D003907:Dexamethasone; D019061:src-Family Kinases