Figure 2
Figure 2. Hematopoietic stem and progenitor-cell populations are preserved in fetal livers of Stat5abnull/null mice. (A) Flow cytometric analysis of Stat5abnull/null, Stat5ab+/null, and Stat5ab+/+ 14.5-dpc fetal livers for early hematopoietic cells. Top panel shows Kit and Sca-1 expression of lineage-negative fetal liver cells. Bottom panel shows CMP (FcγRloCD34+), MEP (FcγRloCD34−), and GMP (FcγRhiCD34+) populations, with percentage of Kit+Lin−Sca− falling into each progenitor population as indicated. (B) Proportion of cells that is identified as either KLS (black), progenitors (CMPs, MEPs, or GMPs, diagonal stripes), or lineage-committed (gray) hematopoietic cells within Stat5ab+/+, Stat5ab+/null, and Stat5abnull/null 14.5-dpc fetal livers as determined by flow cytometric detection of cell-surface markers. (C) Cytokine-dependent colony formation of vector-transduced 14.5-dpc fetal liver cells when cells were plated in the presence of SCF, IL3, IL6, and EPO. Data shown are the mean percentage of colonies formed by Stat5abnull/null and Stat5ab+/null cells relative to Stat5ab+/+ cells. Error bars represent standard deviation. Vector control Stat5ab+/+ cells generated 688 ± 18 G418-resistant colonies per 105 transduced fetal liver cells, while Stat5ab+/null and Stat5abnull/null cells formed 535 ± 12 and 355 ± 10 colonies. TPNeo-transduced Stat5ab+/+, Stat5ab+/null, and Stat5abnull/null cells formed 337 ± 33, 559 ± 43, and 321 ± 30 G418 colonies, respectively. Transduction efficiency of Stat5ab+/+ cells averaged 25% ± 15% and Stat5abnull/null cells averaged 45% ± 16% for 7 independent transductions of each genotype. (KLS indicates Kit+Sca+Lin−; CMP, common myeloid progenitor; MEP, megakaryocyte-erythrocyte progenitor; and GMP, granulocyte-monocyte progenitor.)

Hematopoietic stem and progenitor-cell populations are preserved in fetal livers of Stat5abnull/null mice. (A) Flow cytometric analysis of Stat5abnull/null, Stat5ab+/null, and Stat5ab+/+ 14.5-dpc fetal livers for early hematopoietic cells. Top panel shows Kit and Sca-1 expression of lineage-negative fetal liver cells. Bottom panel shows CMP (FcγRloCD34+), MEP (FcγRloCD34), and GMP (FcγRhiCD34+) populations, with percentage of Kit+LinSca falling into each progenitor population as indicated. (B) Proportion of cells that is identified as either KLS (black), progenitors (CMPs, MEPs, or GMPs, diagonal stripes), or lineage-committed (gray) hematopoietic cells within Stat5ab+/+, Stat5ab+/null, and Stat5abnull/null 14.5-dpc fetal livers as determined by flow cytometric detection of cell-surface markers. (C) Cytokine-dependent colony formation of vector-transduced 14.5-dpc fetal liver cells when cells were plated in the presence of SCF, IL3, IL6, and EPO. Data shown are the mean percentage of colonies formed by Stat5abnull/null and Stat5ab+/null cells relative to Stat5ab+/+ cells. Error bars represent standard deviation. Vector control Stat5ab+/+ cells generated 688 ± 18 G418-resistant colonies per 105 transduced fetal liver cells, while Stat5ab+/null and Stat5abnull/null cells formed 535 ± 12 and 355 ± 10 colonies. TPNeo-transduced Stat5ab+/+, Stat5ab+/null, and Stat5abnull/null cells formed 337 ± 33, 559 ± 43, and 321 ± 30 G418 colonies, respectively. Transduction efficiency of Stat5ab+/+ cells averaged 25% ± 15% and Stat5abnull/null cells averaged 45% ± 16% for 7 independent transductions of each genotype. (KLS indicates Kit+Sca+Lin; CMP, common myeloid progenitor; MEP, megakaryocyte-erythrocyte progenitor; and GMP, granulocyte-monocyte progenitor.)

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