Journal: Cell

Article Title: Mobilization-based chemotherapy-free engraftment of gene-edited human hematopoietic stem cells

doi: 10.1016/j.cell.2022.04.039

Figure Lengend Snippet: M-HSCT allows establishing sufficient donor chimerism to rescue the HIGM1 phenotype (A) Schematic of different mobilization protocols tested and the times of analysis in Cd40lg −/− mice. (B–D) Counts of mobilized WBC (B), LSK (C), and SLAM HSC (D) per mL in the PB of Cd40lg −/− mice treated with PBS (Sham), G-CSF for 7 days (G7), G-CSF for 7 days and AMD3100 (G7A), G-CSF for 7 days, AMD3100 and BIO5192 (G7AB), half-dose of G-CSF for 7 days, AMD3100 and BIO5192 (G7AB-H), G-CSF for 5 days and AMD3100 (G5A), G-CSF for 5 days, AMD3100 and BIO5192 (G5AB), G-CSF for 3 days, AMD3100 and BIO5192 (G3AB), half-dose of G-CSF for 3 days, AMD3100 and BIO5192 (G3AB-H), and only AMD3100 and BIO5192 (AB) at 0, 1, 3, 6, and 9 h after the last injection of A or AB. (E and F) MMP9 (E) and CXCL12 (F) concentration in the BM extracellular extracts of Cd40lg −/− mice mobilized with protocols described above. (G) Total counts of SLAM HSC in the lower limbs (left panel) and in the PB per mL (right panel) of Cd40lg −/− mice treated with PBS or mobilized with G7AB. (H) Long-term follow-up of the donor WT (blue) and recipient Cd40lg −/− (ocher) chimerism observed within total CD45 + cells in PB after transplanting 2 × 10 6 Lin − cells (collected from the BM) post mobilization in recipient Cd40lg −/− mice. (I–K) Myeloid and lymphoid lineage composition of Cd40lg −/− (left panel) and WT (right panel) cells in the PB (I), BM (J), and spleen (K) of Cd40lg −/− mice after M-HSCT. (L) TNP-KLH-specific IgG concentration in sera collected 7 days before (pre) and after (post) TNP-KLH vaccination of Cd40lg −/− mice after M-HSCT. (M) The percentage of PNA + GL7 + splenic germinal centers B cells within the spleen of Cd40lg −/− mice after TNP-KLH vaccination of Cd40lg −/− mice treated by M-HSCT. The results are mean ± SEM, with n ≧ 5 biological replicates for the kinetic experiments, except for the AB group (n = 4), and with n ≧ 9 biological replicates for the Cd40lg −/− M-HSCT experiments. In all the analyses, p less than 0.05 were considered significant (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. “ns” means non-significance).

Article Snippet: Mouse SDF-1 alpha / CXCL12 alpha ELISA Kit, for serum, plasma, and cell culture , Sigma-Aldrich , Cat# RAB0125-1KT.

Techniques: Injection, Concentration Assay

Journal: Cell

Article Title: Mobilization-based chemotherapy-free engraftment of gene-edited human hematopoietic stem cells

doi: 10.1016/j.cell.2022.04.039

Figure Lengend Snippet:

Article Snippet: Mouse SDF-1 alpha / CXCL12 alpha ELISA Kit, for serum, plasma, and cell culture , Sigma-Aldrich , Cat# RAB0125-1KT.

Techniques: Blocking Assay, Plasmid Preparation, Recombinant, Electroporation, CRISPR, Enzyme-linked Immunosorbent Assay, Cell Culture, Expressing