Novel culture system of mesenchymal stromal cells from human subcutaneous adipose tissue.
Accumulating evidence suggests that the delivery of human adipose tissue-derived stromal cells (hASCs) has great potential as regenerative therapy. This was performed to develop a method for expanding hASCs by reducing the amount of serum required. We demonstrate that hASCs were able to expand efficiently in media containing 2% serum and fibroblast growth factor-2. These cells, or low serum cultured hASCs (hLASCs), expressed cell surface markers similar to those on bone marrow-derived mesenchymal stem cells, and could be differentiated into cells of mesenchymal lineage. Of interest, hLASCs secreted higher levels of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) than hASCs cultured in 20% serum (hHASCs). Moreover, hLASC-conditioned media significantly increased endothelial cell (EC) proliferation and decreased EC apoptosis compared to that obtained from hHASCs or control media only. Antibodies against VEGF and HGF virtually negated these effects. When hASCs were administered into the ischemic hindlimbs of nude rats, hLASCs improved blood flow, increased capillary density, and raised the levels of VEGF and HGF in the muscles as compared with hHASCs. In conclusion, we demonstrate a novel low serum culture system for hASCs, which may have great potential in regenerative cell therapy for damaged organs in the clinical setting.
Iwashima
Shigejiro
S
Department of Nephrology, Nagoya University Graduate School of Medicine, Showaku, Nagoya, Aichi, Japan.
Ozaki
Takenori
T
Maruyama
Shoichi
S
Saka
Yousuke
Y
Kobori
Masato
M
Omae
Kaoru
K
Yamaguchi
Hirotake
H
Niimi
Tomoaki
T
Toriyama
Kazuhiro
K
Kamei
Yuzuru
Y
Torii
Shuhei
S
Murohara
Toyoaki
T
Yuzawa
Yukio
Y
Kitagawa
Yasuo
Y
Matsuo
Seiichi
S
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology; D048430:Cell Shape; D064987:Cell- and Tissue-Based Therapy / Q000379:methods*; D054377:Chemokine CXCL12 / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000737:chemistry; D016207:Cytokines / Q000378:metabolism; D017228:Hepatocyte Growth Factor / Q000378:metabolism; D006614:Hindlimb / Q000098:blood supply; D006801:Humans; D007511:Ischemia / Q000378:metabolism / Q000628:therapy; D017078:Laser-Doppler Flowmetry; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology; D051381:Rats; D011916:Rats, Inbred F344; D011923:Rats, Nude; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology; D050151:Subcutaneous Fat / Q000166:cytology*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
D015415:Biomarkers; D054377:Chemokine CXCL12; D017077:Culture Media, Conditioned; D016207:Cytokines; D042461:Vascular Endothelial Growth Factor A; D017228:Hepatocyte Growth Factor