Human adipose-derived mesenchymal stem cells in vitro: evaluation of an optimal expansion medium preserving stemness.
OBJECTIVE
The potential of cultured adipose-derived stem cells (ASC) in regenerative medicine and new cell therapeutic concepts has been shown recently by many investigations. However, while the method of isolation of ASC from liposuction aspirates depending on plastic adhesion is well established, a standard expansion medium optimally maintaining the undifferentiated state has not been described.


METHODS
We cultured ASC in five commonly used culture media (two laboratory-made media and three commercially available media) and compared them with a standard medium. We analyzed the effects on cell morphology, proliferation, hepatocyte growth factor (HGF) expression, stem cell marker profile and differentiation potential. Proliferation was measured with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and a fluorescent assay. Release of HGF was assessed by an immunoassay. Expression of characteristic stem cell-related transcription factors and markers was evaluated by quantitative polymerase chain reaction (qPCR) (Nanog, Sox-2, Rex-1, nestin and Oct-4) and flow cytometry (CD44, CD73, CD90, CD105 and CD166), and differentiation was shown by adipogenic medium.


RESULTS
The morphology and expansion of ASC were significantly affected by the media used, whereas none of the media influenced the ASC potential to differentiate into adipocytes. Furthermore, two of the media induced an increase in expression of transcription factors, an increased secretion of HGF and a decrease in CD105 expression.


CONCLUSIONS
Culture of ASC in one of these two media before using the cells in cell therapeutic approaches may have a benefit on their regenerative potential.
Baer
Patrick C
PC
Division of Nephrology, Department of Internal Medicine III, Goethe University, Frankfurt, Germany. P.Baer@em.uni-frankfurt.de
Griesche
Nadine
N
Luttmann
Werner
W
Schubert
Ralf
R
Luttmann
Arlette
A
Geiger
Helmut
H
D017667:Adipocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D015703:Antigens, CD / Q000032:analysis / Q000378:metabolism; D000954:Antigens, Surface / Q000032:analysis / Q000378:metabolism; D001681:Biological Assay; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology*; D000071063:Endoglin; D005260:Female; D017228:Hepatocyte Growth Factor / Q000032:analysis / Q000378:metabolism; D006801:Humans; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D011956:Receptors, Cell Surface / Q000032:analysis / Q000378:metabolism; D020127:Recovery of Function / Q000502:physiology; D012038:Regeneration / Q000187:drug effects / Q000502:physiology*; D014157:Transcription Factors / Q000032:analysis / Q000378:metabolism; D015533:Transcriptional Activation / Q000187:drug effects / Q000502:physiology
D015703:Antigens, CD; D000954:Antigens, Surface; D015415:Biomarkers; D003470:Culture Media; C064206:ENG protein, human; D000071063:Endoglin; D011956:Receptors, Cell Surface; D014157:Transcription Factors; D017228:Hepatocyte Growth Factor