Journal: BMC Musculoskeletal Disorders

Article Title: Fluorescence activated enrichment of CD146+ cells during expansion of human bone-marrow derived mesenchymal stromal cells augments proliferation and GAG/DNA content in chondrogenic media

doi: 10.1186/1471-2474-15-322

Figure Lengend Snippet: Surface marker distribution on MSCs. a) MSCs were clearly negative for CD14 and CD34 while CD45 was expressed on some of the cells. MSCs were positive for CD73, CD90 and CD105. The histograms displayed are P2 control MSCs cultured in DMEM-LG. All histograms have been adjusted for height. b) Representative histograms from P0 cells in DMEM and ES medium. CD146 expression was significantly lower in ES than in DMEM in P0. c) Exemplary histograms of P2 control vs. FACS-sorted MSCs. d) Exemplary histograms of P2 control vs. MACS-sorted MSCs. There was an important variation of CD146 expression between patients, as demonstrated by the P2 samples from c and d. Magnetic sorting resulted in a lower purification than fluorescence sorting, while higher CD146 expression in naïve MSCs resulted in a higher purification result.

Article Snippet: After an incubation of 15 min, cells were washed and resuspended in the auto MACS buffer, and magnetic isolation was performed according to manufacturers’ protocols using LS columns and a MidiMACSTM separator (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Marker, Control, Cell Culture, Expressing, Purification, Fluorescence

Journal: BMC Musculoskeletal Disorders

Article Title: Fluorescence activated enrichment of CD146+ cells during expansion of human bone-marrow derived mesenchymal stromal cells augments proliferation and GAG/DNA content in chondrogenic media

doi: 10.1186/1471-2474-15-322

Figure Lengend Snippet: Osteogenic, adipogenic and chondrogenic differentiation results. a) Alizarin red contents per protein at days 3, 7, 14 and 21 are displayed in the right diagram, ALP contents per protein per ml and min at days 3, 7, 14 and 21 are displayed in the left diagram. No significant differences between fluorescence or magnetic sorting and the control MSCs could be detected. b) No differences regarding adipogenic differentiation of MSCs cultured in DMEM or ES medium and sorted for CD146 by fluorescence cytometry (DMEM-FACS and ES-FACS) or magnetically (DMEM-MACS and ES-MACS) could be detected. Light microscopy, magnification x100. c) Mean GAG/DNA content of control MSCs, FACS and MACS sorted cells after 21 days of chondrogenic differentiation is displayed. *p < 0.05.

Article Snippet: After an incubation of 15 min, cells were washed and resuspended in the auto MACS buffer, and magnetic isolation was performed according to manufacturers’ protocols using LS columns and a MidiMACSTM separator (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Fluorescence, Control, Cell Culture, Cytometry, Light Microscopy