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Bone morphogenetic protein 4 induces efficient hematopoietic differentiation of rhesus monkey embryonic stem cells in vitro by Fei Li, Shijiang Lu, Loyda.

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Presentation on theme: "Bone morphogenetic protein 4 induces efficient hematopoietic differentiation of rhesus monkey embryonic stem cells in vitro by Fei Li, Shijiang Lu, Loyda."— Presentation transcript:

1 Bone morphogenetic protein 4 induces efficient hematopoietic differentiation of rhesus monkey embryonic stem cells in vitro by Fei Li, Shijiang Lu, Loyda Vida, James A. Thomson, and George R. Honig Blood Volume 98(2): July 15, 2001 ©2001 by American Society of Hematology

2 Phase-contrast photomicrograph of an undifferentiated rhesus monkey ES cell colony (magnification, ×100). Phase-contrast photomicrograph of an undifferentiated rhesus monkey ES cell colony (magnification, ×100). Fei Li et al. Blood 2001;98: ©2001 by American Society of Hematology

3 Phase-contrast photomicrographs of hematopoietic-like clusters from differentiated ES colonies.(A) A hematopoietic-like cluster (magnification, ×100). Phase-contrast photomicrographs of hematopoietic-like clusters from differentiated ES colonies.(A) A hematopoietic-like cluster (magnification, ×100). (B) A hematopoietic-like cluster showing morphologic characteristics similar to an embryonic blood island (magnification, ×100). (C) A hematopoietic-like cluster induced by BMP-4 (magnification, ×100). Fei Li et al. Blood 2001;98: ©2001 by American Society of Hematology

4 Phase-contrast photomicrographs of expanded hematopoietic-like clusters from differentiated ES colonies.(A) A cluster expanded with cytokines (magnification, ×100). Phase-contrast photomicrographs of expanded hematopoietic-like clusters from differentiated ES colonies.(A) A cluster expanded with cytokines (magnification, ×100). (B) A migrated CAFC in primary differentiation culture (magnification, ×100). Fei Li et al. Blood 2001;98: ©2001 by American Society of Hematology

5 Photomicrographs of hematopoietic progeny cells derived from differentiated rhesus monkey ES colonies.(A) Granulocyte and monocyte. Photomicrographs of hematopoietic progeny cells derived from differentiated rhesus monkey ES colonies.(A) Granulocyte and monocyte. (B) Macrophage. (C,D) Granulocytes. (E) Megakaryocyte. (F,G) Immature blast cells. (H,I) Erythroblasts. Fei Li et al. Blood 2001;98: ©2001 by American Society of Hematology

6 Phase-contrast photomicrographs of replated CAFC colonies from differentiated ES cells.(A) Secondary CAFC colony (magnification, ×100). Phase-contrast photomicrographs of replated CAFC colonies from differentiated ES cells.(A) Secondary CAFC colony (magnification, ×100). (B) Tertiary CAFC colony (magnification, ×100). Fei Li et al. Blood 2001;98: ©2001 by American Society of Hematology

7 Phase-contrast photomicrographs of CD34+ cells from differentiated ES cell colonies.(A,B) In situ immunofluorescence staining of CD34+ cells (magnification, ×100). Phase-contrast photomicrographs of CD34+ cells from differentiated ES cell colonies.(A,B) In situ immunofluorescence staining of CD34+ cells (magnification, ×100). (C) Immunofluorescence staining of CD34+ cells from a cytospin preparation (magnification, ×200). Fei Li et al. Blood 2001;98: ©2001 by American Society of Hematology

8 Immunofluorescence flow cytometry analysis of CD34 surface markers of differentiated rhesus monkey ES cells.CD34+ cells were defined as cells with dim and bright fluorescence intensity compared with the isotype control. Immunofluorescence flow cytometry analysis of CD34 surface markers of differentiated rhesus monkey ES cells.CD34+ cells were defined as cells with dim and bright fluorescence intensity compared with the isotype control. Fei Li et al. Blood 2001;98: ©2001 by American Society of Hematology

9 Photomicrograph of sorted CD34+ cells from differentiated rhesus monkey ES colonies.Cell populations of medium and small size were gated. Photomicrograph of sorted CD34+ cells from differentiated rhesus monkey ES colonies.Cell populations of medium and small size were gated. CD34+cells with bright fluorescence intensity were sorted and stained with Wright-Giemsa stain after cytospin preparation (magnification, ×1000). Fei Li et al. Blood 2001;98: ©2001 by American Society of Hematology

10 RT-PCR analysis of gene expression by BMP-4–treated ES cells
RT-PCR analysis of gene expression by BMP-4–treated ES cells.BM indicates rhesus monkey bone marrow cells; ES, undifferentiated ES cells; c, BMP-4–treated S17 cells; plus sign, BMP-4–treated ES cells; and minus sign, ES cells without BMP-4. RT-PCR analysis of gene expression by BMP-4–treated ES cells.BM indicates rhesus monkey bone marrow cells; ES, undifferentiated ES cells; c, BMP-4–treated S17 cells; plus sign, BMP-4–treated ES cells; and minus sign, ES cells without BMP-4. Fei Li et al. Blood 2001;98: ©2001 by American Society of Hematology


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