Phenotypic Characterization of ASCs

Freshly isolated cells or ASCs at the third passage in culture were detached by Tryple Express and were incubated with the properly conjugated antibodies CD10, CD13, CD29, CD34, CD44, CD45, CD49a, CD49b, CD49d, CD59 (BD Biosciences, Milan, Italy), CD66e (AbD Serotec, Kidlington, UK), CD73, CD90 (BD Biosciences), CD105 (Serotec), CD117 (BD Biosciences), CD133 (Miltenyi Biotec, Bergisch Gladbach, Germany), CD271 (R&D Systems, Minneapolis, MN, USA), HLA-DR (BD Biosciences), KDR and ABCG-2 (R&D System). Properly conjugated isotype-matched antibodies were used as a negative control. The analysis was performed either by FACS-Calibur or by FACSCanto (BD Bioscience) [6 (link), 7 (link), 9 (link), 27 (link)]. Erythrocytes, in freshly isolated samples, were lysed by incubating cells for 5 minutes in fluorescence-activated cell sorting lysing solution (BD Bioscience).

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Domenis R., Lazzaro L., Calabrese S., Mangoni D., Gallelli A., Bourkoula E., Manini I., Bergamin N., Toffoletto B., Beltrami C.A., Beltrami A.P., Cesselli D, & Parodi P.C. (2015). Adipose tissue derived stem cells: in vitro and in vivo analysis of a standard and three commercially available cell-assisted lipotransfer techniques. Stem Cell Research & Therapy, 6(1), 2.

Publication 2015

CD49a CD49b CD49d CD105 CD117 CD133 CD271 HLA-DR ABCG-2 FACS-Calibur FACSCanto fluorescence-activated cell sorting lysing solution

Antibodies Ascs Cd271 Cd44 Cd59 Cd73 Cd90 Cells Erythrocytes Hla dr Isotype

Corresponding Organization : University of Udine

Other organizations : Ospedale Santa Maria della Misericordia di Udine

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Variable analysis

independent variables

Passage number (third passage)

dependent variables

Expression of surface markers CD10, CD13, CD29, CD34, CD44, CD45, CD49a, CD49b, CD49d, CD59, CD66e, CD73, CD90, CD105, CD117, CD133, CD271, HLA-DR, KDR, and ABCG-2

control variables

Cell type (freshly isolated cells or ASCs)
Cell detachment method (Tryple Express)
Antibodies used for staining (properly conjugated)
Isotype-matched antibodies used as negative control
Flow cytometry instrument used (FACS-Calibur or FACSCanto)
Erythrocyte lysis (fluorescence-activated cell sorting lysing solution)

positive controls

Isotype-matched antibodies

negative controls

Isotype-matched antibodies

Annotations

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