DOI: 10.6084/m9.figshare.13467161
title: Additional file 1 of Quantitative assessment of the impact of cryopreservation on human bone marrow-derived mesenchymal stem cells: up to 24 h post-thaw and beyond

description: Additional file 1: Figures S1-S8. Representative histograms for immunophenotyping, viability and apoptosis measures for the three lines (fresh and at 0h, 2h, 4h and 24h post-thaw). Figure S1. Immunophenotyping of fresh cells: Representative histograms of expression of CD90, CD105, CD73 and CD14, CD20, CD34, CD45 and HLA-DR all linked to PerCP. Black peaks represent isotype controls in all histograms. The first row of histograms is for M4 (red), the second for M6 (blue) and the third for M7 (green). All measurements were done in triplicates from three independent experiments based on at least 100,000 events. Figure S2. Immunophenotyping of cryopreserved cells at 0 h post-thaw: Representative histograms of expression of CD90, CD105, CD73 and CD14, CD20, CD34, CD45 and HLA-DR all linked to PerCP. Black peaks represent isotype controls in all histograms. The first row of histograms is for M4 (red), the second for M6 (blue) and the third for M7 (green). All measurements were done in triplicates from three independent experiments based on at least 100,000 events. Figure S3. Immunophenotyping of cryopreserved cells at 2 h post-thaw: Representative histograms of expression of CD90, CD105, CD73 and CD14, CD20, CD34, CD45 and HLA-DR all linked to PerCP. Black peaks represent isotype controls in all histograms. The first row of histograms is for M4 (red), the second for M6 (blue) and the third for M7 (green). All measurements were done in triplicates from three independent experiments based on at least 100,000 events. Figure S4. Immunophenotyping of cryopreserved cells at 4 h post-thaw: Representative histograms of expression of CD90, CD105, CD73 and CD14, CD20, CD34, CD45 and HLA-DR all linked to PerCP. Black peaks represent isotype controls in all histograms. The first row of histograms is for M4 (red), the second for M6 (blue) and the third for M7 (green). All measurements were done in triplicates from three independent experiments based on at least 100,000 events. Figure S5. Immunophenotyping of cryopreserved cells at 24 h post-thaw: Representative histograms of expression of CD90, CD105, CD73 and CD14, CD20, CD34, CD45 and HLA-DR all linked to PerCP. Black peaks represent isotype controls in all histograms. The first row of histograms is for M4 (red), the second for M6 (blue) and the third for M7 (green). All measurements were done in triplicates from three independent experiments based on at least 100,000 events. Figure S6. Apoptosis and viability measures of the M4 cell line: Representative overlay histograms of blank samples (black) and M4 samples stained with Annexin V and 7AAD (red) at different time points. All measurements were done in triplicates from three independent experiments based on at least 100,000 events. Figure S7. Apoptosis and viability measures of the M6 cell line: Representative overlay histograms of blank samples (black) and M6 samples stained with Annexin V and 7AAD (red) at different time points. All measurements were done in triplicates from three independent experiments based on at least 100,000 events. Figure S8. Apoptosis and viability measures of the M7 cell line: Representative overlay histograms of blank samples (black) and M7 samples stained with Annexin V and 7AAD (green) at different time points. All measurements were done in triplicates from three independent experiments based on at least 100,000 events.