Journal: iScience

Article Title: Epidermal growth factor augments the self-renewal capacity of aged hematopoietic stem cells

doi: 10.1016/j.isci.2024.110306

Figure Lengend Snippet: EGF treatment in vitro improves the competitive repopulating capacity of aged HSCs (A) Total cell counts, %KSL cells and % SLAM KSL cells at day +7 of culture of aged BM 34 − KSL cells with Media alone (TSF) or media + EGF (n = 9–10/group, t test). (B) Schematic representation of the competitive transplantation assays performed. Recipient mice irradiated with 875 cGy TBI. (C) Donor CD45.2+ cell engraftment over time in the PB of primary recipient CD45.1+ mice transplanted competitively with the progeny of 500 aged BM 34-KSL cells cultured with media alone or media + EGF x 7 days (n = 9-10/group). (D) Representative flow cytometric analysis of donor CD45.2 + cell engraftment in the PB of CD45.1 + recipient mice at 16 weeks following transplantation as described in (C). (E) Mean percentages of donor CD45.2 + , CD45.2 + Mac-1/Gr-1 + cells, CD45.2 + B220 + cells, and CD45.2 + CD3 + cells in the PB of CD45.1 + recipient mice at 16 weeks post competitive transplantation of the progeny of 500 aged BM 34 − KSL cells treated with media alone or media + EGF ( n = 9–10/group, ANOVA). (F) Representative flow cytometric analysis of donor CD45.2 + cell engraftment within the BM KSL population and within BM CD150 + CD48 − KSL HSCs at week 16 in recipient mice transplanted with the progeny of aged BM 34 − KSL cells treated with media alone or media + EGF. (G) Mean %CD45.2 + KSL cells and %CD45.2 + CD150 + CD48 − KSL (SLAM KSL) cells in recipient mice transplanted with the progeny of culture with media alone or media + EGF ( n = 9/group, t test). (H) Mean percentages of donor CD45.2 + cells in the PB of secondary mice that were transplanted with 1 × 10 6 BM cells collected at 16 weeks from primary recipient mice that were competitively transplanted with the progeny of aged BM 34 − KSL cells cultured × 7 days with media alone or media + EGF ( n = 9/group, t test). Data are represented as means +/− SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: APC-Cy7 B220 (CD45R) , BD Biosciences , 552094.

Techniques: In Vitro, Transplantation Assay, Irradiation, Cell Culture

Journal: iScience

Article Title: Epidermal growth factor augments the self-renewal capacity of aged hematopoietic stem cells

doi: 10.1016/j.isci.2024.110306

Figure Lengend Snippet:

Article Snippet: APC-Cy7 B220 (CD45R) , BD Biosciences , 552094.

Techniques: Purification, Recombinant, Enzyme-linked Immunosorbent Assay, Reverse Transcription, Sequencing, Software

Journal: iScience

Article Title: Epidermal growth factor augments the self-renewal capacity of aged hematopoietic stem cells

doi: 10.1016/j.isci.2024.110306

Figure Lengend Snippet: EGF treatment in vitro improves the competitive repopulating capacity of aged HSCs (A) Total cell counts, %KSL cells and % SLAM KSL cells at day +7 of culture of aged BM 34 − KSL cells with Media alone (TSF) or media + EGF (n = 9–10/group, t test). (B) Schematic representation of the competitive transplantation assays performed. Recipient mice irradiated with 875 cGy TBI. (C) Donor CD45.2+ cell engraftment over time in the PB of primary recipient CD45.1+ mice transplanted competitively with the progeny of 500 aged BM 34-KSL cells cultured with media alone or media + EGF x 7 days (n = 9-10/group). (D) Representative flow cytometric analysis of donor CD45.2 + cell engraftment in the PB of CD45.1 + recipient mice at 16 weeks following transplantation as described in (C). (E) Mean percentages of donor CD45.2 + , CD45.2 + Mac-1/Gr-1 + cells, CD45.2 + B220 + cells, and CD45.2 + CD3 + cells in the PB of CD45.1 + recipient mice at 16 weeks post competitive transplantation of the progeny of 500 aged BM 34 − KSL cells treated with media alone or media + EGF ( n = 9–10/group, ANOVA). (F) Representative flow cytometric analysis of donor CD45.2 + cell engraftment within the BM KSL population and within BM CD150 + CD48 − KSL HSCs at week 16 in recipient mice transplanted with the progeny of aged BM 34 − KSL cells treated with media alone or media + EGF. (G) Mean %CD45.2 + KSL cells and %CD45.2 + CD150 + CD48 − KSL (SLAM KSL) cells in recipient mice transplanted with the progeny of culture with media alone or media + EGF ( n = 9/group, t test). (H) Mean percentages of donor CD45.2 + cells in the PB of secondary mice that were transplanted with 1 × 10 6 BM cells collected at 16 weeks from primary recipient mice that were competitively transplanted with the progeny of aged BM 34 − KSL cells cultured × 7 days with media alone or media + EGF ( n = 9/group, t test). Data are represented as means +/− SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Lineage analysis was performed using PE Mac-1 (CD11b) (Cat # 557397), PE Gr-1 (Ly-6G and Ly-6C) (553128), APC Ter-119 (557909), V450 CD3 (561389), APC-Cy7 B220 (CD45R) (552094) antibodies purchased from BD Biosciences, San Jose, CA.

Techniques: In Vitro, Transplantation Assay, Irradiation, Cell Culture

Journal: iScience

Article Title: Epidermal growth factor augments the self-renewal capacity of aged hematopoietic stem cells

doi: 10.1016/j.isci.2024.110306

Figure Lengend Snippet:

Article Snippet: Lineage analysis was performed using PE Mac-1 (CD11b) (Cat # 557397), PE Gr-1 (Ly-6G and Ly-6C) (553128), APC Ter-119 (557909), V450 CD3 (561389), APC-Cy7 B220 (CD45R) (552094) antibodies purchased from BD Biosciences, San Jose, CA.

Techniques: Purification, Recombinant, Enzyme-linked Immunosorbent Assay, Reverse Transcription, Sequencing, Software