Figure 5. Cultured HSCs can enhance early hematopoietic recovery. (A) Photomicrographs of cultured HSCs at indicated times. Photos were taken and recorded by NIKON digital sight DS-Fi1, L2, and U2 (NIKON) through a microscope (NIKON ECLIPSE TS100, ocular lenses 10X, objective lenses 4X NA 0.13, W.D 16.5). Bars, 300 µm. (B) Numbers of cultured cells in total or of each indicated population in a well counted by flow cytometry (input = CD34−KSL: 50 cells). One-way-ANOVA with Tukey’s multiple comparison test was used for statistical analysis. n = 5 in each group. Mean values are indicated as bars. P < 0.05 was considered statistically significant. *, P < 0.05; ***, P < 0 0.001; ****, P < 0 0.0001. (C) The experimental protocol to test bridging effects in cultured HSCs. Cultured or uncultured HSCs (CD34− c-Kit+ Sca1+ Lin− cells) derived from B6 (Ly5.1) mice were competitively transplanted with uncultured B6 (Ly5.1/5.2) WBM cells into lethally irradiated B6 (Ly5.2) mice. (D) Donor cell (B6-Ly5.1) contribution to a PB myeloid lineage (granulocytes and monocytes) assessed at indicated times. n = 6 in each group. Mann-Whitney test was used for statistical analysis. Mean values are indicated as bars. P < 0.05 was considered statistically significant. ****, P < 0 0.0001. (E, left) Phenotypic characteristics of 7-d cultured CD34−KSL cells. Note that these cells in total are mostly Lin−c-Kit+ (∼97–98%). Listed subfractions (CD48+ Sca1+ Lin− c-Kit+, 12.8%; CD48+ Sca1− Lin− c-Kit+, 81.2%; CD48− Sca1+ Lin− c-Kit+, 5.2%) derived from B6 (Ly5.1) were competitively transplanted with 106 WBM cells from B6 (Ly5.1/5.2) into lethally irradiated B6 (Ly5.2) mice. (right) Donor cell (B6-Ly5.1) contribution to a PB myeloid lineage (granulocytes and monocytes) assessed at indicated times. n = 8 in each group. (F) Donor cell contributions in RBCs and platelets detected by KuO fluorescence at indicated times. n = 6 in each group. Mann-Whitney testing was used for statistical analysis. Mean values are indicated as bars. P < 0.05 was considered statistically significant. *, P < 0.05; **, P < 0.01. Representative data are shown from at least two independent experiments for each panel.
Figure 5.

Cultured HSCs can enhance early hematopoietic recovery. (A) Photomicrographs of cultured HSCs at indicated times. Photos were taken and recorded by NIKON digital sight DS-Fi1, L2, and U2 (NIKON) through a microscope (NIKON ECLIPSE TS100, ocular lenses 10X, objective lenses 4X NA 0.13, W.D 16.5). Bars, 300 µm. (B) Numbers of cultured cells in total or of each indicated population in a well counted by flow cytometry (input = CD34−KSL: 50 cells). One-way-ANOVA with Tukey’s multiple comparison test was used for statistical analysis. n = 5 in each group. Mean values are indicated as bars. P < 0.05 was considered statistically significant. *, P < 0.05; ***, P < 0 0.001; ****, P < 0 0.0001. (C) The experimental protocol to test bridging effects in cultured HSCs. Cultured or uncultured HSCs (CD34− c-Kit+ Sca1+ Lin− cells) derived from B6 (Ly5.1) mice were competitively transplanted with uncultured B6 (Ly5.1/5.2) WBM cells into lethally irradiated B6 (Ly5.2) mice. (D) Donor cell (B6-Ly5.1) contribution to a PB myeloid lineage (granulocytes and monocytes) assessed at indicated times. n = 6 in each group. Mann-Whitney test was used for statistical analysis. Mean values are indicated as bars. P < 0.05 was considered statistically significant. ****, P < 0 0.0001. (E, left) Phenotypic characteristics of 7-d cultured CD34−KSL cells. Note that these cells in total are mostly Lin−c-Kit+ (∼97–98%). Listed subfractions (CD48+ Sca1+ Lin− c-Kit+, 12.8%; CD48+ Sca1− Lin− c-Kit+, 81.2%; CD48− Sca1+ Lin− c-Kit+, 5.2%) derived from B6 (Ly5.1) were competitively transplanted with 106 WBM cells from B6 (Ly5.1/5.2) into lethally irradiated B6 (Ly5.2) mice. (right) Donor cell (B6-Ly5.1) contribution to a PB myeloid lineage (granulocytes and monocytes) assessed at indicated times. n = 8 in each group. (F) Donor cell contributions in RBCs and platelets detected by KuO fluorescence at indicated times. n = 6 in each group. Mann-Whitney testing was used for statistical analysis. Mean values are indicated as bars. P < 0.05 was considered statistically significant. *, P < 0.05; **, P < 0.01. Representative data are shown from at least two independent experiments for each panel.

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