Figure 5
Cellular and molecular properties of MSC cultures established by Protocol 2.
(a) Phase contrast images taken after 24 h of initial plating (P0, D1), 10 days (P0, D10) and on day 3 of passage 5 (P5, D3) are shown. (b) FISH images of adherent cell cultures at day 3 of passage 1 showing two X chromosomes-specific signals in all cells (a-b, scale bar, 10 μm). A similar FISH analysis was also done at passage 5 (Supplementary Figure S9). (c) Electropherograms showing the STR profiles of the mother and the adherent cell cultures (MSC) at passage 1. The STR profile of the MSC perfectly matched with the STR profile of the mother (maternal blood). (d) Immunofluorescence images of maternal cells at passage 1 showing low or no fluorescence for E-cadherin and CK18 markers (scale bars, 50 μm). Image of the antibody negative control is shown in Supplementary Figure S5. (e) Percentage of cells expressing MSC-specific surface markers (first 6 columns) as calculated by flow cytometry (mean ± standard deviation, n = 3). Histograms for each marker from a representative experiment are shown in Supplementary Figure S10. (f) Tri-lineage differentiation of MSC at passage 5 - adipocytes (Oil red O staining), osteoblasts (von Kossa staining) and chondrocytes (Alcian blue staining) (scale bars, 100 μm). (g) GTG-banded chromosome analysis of maternal MSC at passage 5 exhibiting a normal 46, XX karyotype. Corresponding metaphase spread for the karyotype is shown in Supplementary Figure S11.
