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Xenogen-free isolation and culture of human adipose mesenchymal stem cells

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Abstract
Background: Adipose-derived Stem Cells (ASCs) present great potential for reconstructive procedures. Currently, isolation by enzyme digestion and culturing using xenogenic substances remain the gold standard, impairing clinical use. Methods: Abdominal lipo-aspirate and blood samples were obtained from healthy patients. A novel mechanical isolation method for ASCs was compared to (the standard) collagenase digestion. ASCs are examined by flow-cytometry and multilineage differentiation assays. Cell cultures were performed without xenogenic or toxic substances, using autologous plasma extracted from peripheral blood. After eGFP-transfection, an in vivo differentiation assay was performed. Results: Mechanical isolation is more successful in isolating CD34(+)/CD31(-)/CD45(-)/CD13(+)/CD73(+)/CD146(-) ASCs from lipo-aspirate than isolation via collagenase digestion (p < 0.05). ASCs display multilineage differentiation potential in vitro. Autologous plasma is a valid additive for ASCs culturing. eGFP-ASCs, retrieved after 3 months in vivo, differentiated in adipocytes and endothelial cells. Conclusion: A practical method for human ASC isolation and culturing from abdominal lipo-aspirate, without the addition of xenogenic substances, is described. The mechanical protocol is more successful than the current gold standard protocol of enzyme digestion. These results are important in the translation of laboratory-based cell cultures to clinical reconstructive and aesthetic applications.
Keywords
STROMAL-VASCULAR FRACTION, SERUM-FREE CULTURE, BONE-MARROW, FAT GRAFTS, TISSUE, COLLAGENASE, SURVIVAL, Adipose-derived stem cells, Mechanical isolation, Xenogen-free, Autologous plasma

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MLA
Doornaert, Maarten, et al. “Xenogen-Free Isolation and Culture of Human Adipose Mesenchymal Stem Cells.” STEM CELL RESEARCH, vol. 40, 2019, doi:10.1016/j.scr.2019.101532.
APA
Doornaert, M., De Maere, E., Colle, J., Declercq, H., Taminau, J., Lemeire, K., … Blondeel, P. (2019). Xenogen-free isolation and culture of human adipose mesenchymal stem cells. STEM CELL RESEARCH, 40. https://doi.org/10.1016/j.scr.2019.101532
Chicago author-date
Doornaert, Maarten, Elisabeth De Maere, Julien Colle, Heidi Declercq, Joachim Taminau, Kelly Lemeire, Geert Berx, and Phillip Blondeel. 2019. “Xenogen-Free Isolation and Culture of Human Adipose Mesenchymal Stem Cells.” STEM CELL RESEARCH 40. https://doi.org/10.1016/j.scr.2019.101532.
Chicago author-date (all authors)
Doornaert, Maarten, Elisabeth De Maere, Julien Colle, Heidi Declercq, Joachim Taminau, Kelly Lemeire, Geert Berx, and Phillip Blondeel. 2019. “Xenogen-Free Isolation and Culture of Human Adipose Mesenchymal Stem Cells.” STEM CELL RESEARCH 40. doi:10.1016/j.scr.2019.101532.
Vancouver
1.
Doornaert M, De Maere E, Colle J, Declercq H, Taminau J, Lemeire K, et al. Xenogen-free isolation and culture of human adipose mesenchymal stem cells. STEM CELL RESEARCH. 2019;40.
IEEE
[1]
M. Doornaert et al., “Xenogen-free isolation and culture of human adipose mesenchymal stem cells,” STEM CELL RESEARCH, vol. 40, 2019.
@article{8634341,
  abstract     = {{Background: Adipose-derived Stem Cells (ASCs) present great potential for reconstructive procedures. Currently, isolation by enzyme digestion and culturing using xenogenic substances remain the gold standard, impairing clinical use. 
Methods: Abdominal lipo-aspirate and blood samples were obtained from healthy patients. A novel mechanical isolation method for ASCs was compared to (the standard) collagenase digestion. ASCs are examined by flow-cytometry and multilineage differentiation assays. Cell cultures were performed without xenogenic or toxic substances, using autologous plasma extracted from peripheral blood. After eGFP-transfection, an in vivo differentiation assay was performed. 
Results: Mechanical isolation is more successful in isolating CD34(+)/CD31(-)/CD45(-)/CD13(+)/CD73(+)/CD146(-) ASCs from lipo-aspirate than isolation via collagenase digestion (p < 0.05). ASCs display multilineage differentiation potential in vitro. Autologous plasma is a valid additive for ASCs culturing. eGFP-ASCs, retrieved after 3 months in vivo, differentiated in adipocytes and endothelial cells. 
Conclusion: A practical method for human ASC isolation and culturing from abdominal lipo-aspirate, without the addition of xenogenic substances, is described. The mechanical protocol is more successful than the current gold standard protocol of enzyme digestion. These results are important in the translation of laboratory-based cell cultures to clinical reconstructive and aesthetic applications.}},
  articleno    = {{101532}},
  author       = {{Doornaert, Maarten and De Maere, Elisabeth and Colle, Julien and Declercq, Heidi and Taminau, Joachim and Lemeire, Kelly and Berx, Geert and Blondeel, Phillip}},
  issn         = {{1873-5061}},
  journal      = {{STEM CELL RESEARCH}},
  keywords     = {{STROMAL-VASCULAR FRACTION,SERUM-FREE CULTURE,BONE-MARROW,FAT GRAFTS,TISSUE,COLLAGENASE,SURVIVAL,Adipose-derived stem cells,Mechanical isolation,Xenogen-free,Autologous plasma}},
  language     = {{eng}},
  pages        = {{8}},
  title        = {{Xenogen-free isolation and culture of human adipose mesenchymal stem cells}},
  url          = {{http://dx.doi.org/10.1016/j.scr.2019.101532}},
  volume       = {{40}},
  year         = {{2019}},
}

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