Additional file 1 of Small volume bone marrow aspirates with high progenitor cell concentrations maximize cell therapy dose manufacture and substantially reduce donor hemoglobin loss
Creators: name: Epah, Jeremy
name: Spohn, Gabriele
name: Preiß, Kathrin
name: Müller, Markus M.
name: Dörr, Johanna
name: Bauer, Rainer
name: Daqiq-Mirdad, Shabnam
name: Schwäble, Joachim
name: Bernas, Stefanie N.
name: Schmidt, Alexander H.
name: Seifried, Erhard
name: Schäfer, Richard
Publisher: name: figshare
Year: 2023
Subjects: subject: Biophysics
subject: Space Science
subject: Medicine
subject: Cell Biology
subject: Physiology
subject: FOS: Biological sciences
subject: Immunology
subject: FOS: Clinical medicine
subject: Developmental Biology
subject: Cancer
subject: Hematology

description: Additional file 1: Fig. S1. Sampling overview. For each BM collection three small volume BM aspirates (Punctions A, C and E) as well as three large volume BM aspirates (Punctions B, D and F) were sampled at three time points. To standardize sampling and to exclude possible impact of the needle position in the punction canal each sample was taken from separate punctions as first aspirate after reaching the BM cavity. N = 20 donors. Fig. S2. Benchmarking HPC-M collection centers worldwide. A. TNC absolute numbers in HPC-M collected in our center (Center) and in other international collection centers. B. Ratio collected to requested TNC numbers in HPC-M collected in our center and in other international collection centers. One-way ANOVA (Kruskall-Wallis test) followed by Dunn’s multiple comparisons test; error bars: 5 - 95 percentile; N = 81 for our center (Center), N = 829 for GER (Germany without our center), N= 96 for USA, N = 33 for UK, N = 209 for PL.  Fig. S3. Hypothesis. Visualization of the hypothesis that small volume BM aspirates are higher concentrated for progenitor cells compared to large volume BM aspirates that contain more red blood cells thus collecting fewer progenitor cells per milliliter. The main defining factor for the cell types being differentially collected is their specific gravity. Fig. S4. Neutrophils and platelets in small volume aspirates vs. large volume aspirates. A. Neutrophils per mL BM in small volume aspirates vs. large volume aspirates. B. Platelets per mL BM in small volume aspirates vs. large volume aspirates. Wilcoxon matched pairs test; dots: individual samples; horizontal lines: means; error bars: SEM; 20 donors, 1-3 time points, 2 volumes each; small volume samples: N = 47 (neutrophils), N = 57 (platelets); large volume samples: N = 46 (neutrophils), N = 58 (platelets). Fig. S5. Mature CD45pos immune cells in small volume aspirates vs. large volume aspirates. Wilcoxon matched pairs test; dots: individual samples; horizontal lines: means; error bars: SEM; 20 donors, 1-3 time points, 2 volumes each; small volume samples: N = 46; large volume samples: N = 44. Fig. S6. Kinetics of HSPC collection in small volume aspirates vs. large volume aspirates. A. CD34pos HSPCs per mL BM in small volume aspirates vs. large volume aspirates at different time points. Wilcoxon matched pairs test; dots: individual samples; horizontal lines: means; error bars: SEM; t1: small volume samples: N = 14 donors, large volume samples: N = 15 donors; t2: small volume samples: N = 15 donors, large volume samples: N = 15 donors; t3: small volume samples: N= 17 donors, large volume samples: N = 16 donors. B. Time course of CD34pos HSPCs per mL BM in small volume aspirates during BM collection. ANOVA followed by Tukey's multiple comparisons test; dots: means; error bars: SEM; t1: N = 14 donors; t2: N = 15 donors; t3: N = 17 donors. C. Time course of CD34pos HSPCs per mL BM in large volume aspirates during BM collection. ANOVA followed by Tukey's multiple comparisons test; dots: means; error bars: SEM; t1: N = 15 donors t2: N = 15 donors t3: N= 16 donors. Fig. S7. CD45neg MSC subsets in small volume aspirates vs. large volume aspirates. Wilcoxon matched pairs test; dots: individual samples; horizontal lines: means; error bars: SEM; 19 donors, 1-3 time points, 2 volumes each; small volume samples: N = 44; large volume samples: N = 39. Fig. S8. CD45dim MSC subsets in small volume aspirates vs. large volume aspirates. Wilcoxon matched pairs test; dots: individual samples; horizontal lines: means; error bars: SEM; 19 donors, 1-3 time points, 2 volumes each; small volume samples: N = 44; large volume samples: N = 39. Fig. S9. Endothelial progenitor cell subsets in small volume aspirates vs. large volume aspirates. Wilcoxon matched pairs test; dots: individual samples; horizontal lines: means; error bars: SEM; all cells were positive for CD31 and negative for CD45 and CD14; right Y-axis refers to ECFCs, CACs and EPCs; 20 donors, 1-3 time points, 2 volumes each; small volume samples: N = 47; large volume samples: N = 46. Fig. S10. Correlation of TNC count ratios to hematopoietic subsets count ratios at different time points. Red lines: TNC count ratios (small volume to large volume aspirates). Black lines: Hematopoietic subsets count ratios (small volume to large volume aspirates). Position of the regression line above the TNC line indicates a higher cell concentration compared to TNCs in the small volume aspirates and vice versa. Individual data points are only shown for hematopoietic subset ratios. Spearman Correlation: N= 37-41. Fig. S11. Correlation of TNC count ratios to CFU count ratios at different time points. Red lines: TNC count ratios (small volume to large volume aspirates). Black lines: CFU count ratios (small volume to large volume aspirates). Position of the regression line above the TNC line indicates a higher cell concentration compared to TNCs in the small volume aspirates and vice versa. Individual data points are only shown for CFU count ratios. Spearman Correlation: N = 33-41. Fig. S12. Correlation of TNC count ratios to EPC count ratios at different time points. Red lines: TNC count ratios (small volume to large volume aspirates). Black lines: EPC count ratios (small volume to large volume aspirates). Position of the regression line above the TNC line indicates a higher cell concentration compared to TNCs in the small volume aspirates and vice versa. Individual data points are only shown for EPC count ratios. Spearman Correlation: CACs: CD45negCD14negCD31posCD34posCD133pos. EPCs: CD45negCD14negCD31posCD34posCD133posCD309pos. ECFCs: CD45negCD14negCD31posCD34posCD133neg. N = 30-41.