Macrophage Differentiation from Embryoid Bodies Derived from Human Embryonic Stem Cells

Overview

Journal J Stem Cells

Specialty Cell Biology

Date 2010 May 26

PMID 20498689

Citations 12

Authors

Aparna Subramanian

Beichu Guo

Matthew D Marsden

Zoran Galic

Scott Kitchen

Amelia Kacena

Helen J Brown

Genhong Cheng

Jerome A Zack

Affiliations

Soon will be listed here.

Abstract

Human embryonic stem cells can differentiate into CD34+ hematopoietic progenitors by co-culture on murine feeders such as OP9 and S17. These CD34+ progenitors can be further differentiated into several cells of the hematopoietic lineage including macrophages. However, co-culture on murine feeders is time consuming and involves extensive manipulations. Furthermore, CD45 expression is low on hematopoietic cultures derived from stromal co-cultures. In this study we describe a novel and highly efficient system of generating differentiated macrophages from hematopoietic progenitors generated from embryoid body cultures of human embryonic stem cells. The hematopoietic progenitors generated from these embryoid bodies express higher numbers of CD45+ cells and are able to differentiate to macrophages when cultured in presence of cytokines. Using this system we were able to generate higher yields of CD14+ macrophages compared to traditional stromal cell culture methods. The embryoid body derived macrophages are phagocytic, respond to Toll-like receptor stimulation and express phenotypic markers of mature macrophages. Importantly, the embryoid body system generates hematopoietic progenitors suitable for clinical use by eliminating the need for murine feeder cells. Furthermore, this system is amenable to genetic manipulation and may thus be used to study important mechanisms of macrophage differentiation and function.

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References

Ogawa M . Differentiation and proliferation of hematopoietic stem cells. Blood. 1993; 81(11):2844-53. View

Granelli-Piperno A, Pritsker A, Pack M, Shimeliovich I, Arrighi J, Park C . Dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin/CD209 is abundant on macrophages in the normal human lymph node and is not required for dendritic cell stimulation of the mixed leukocyte reaction. J Immunol. 2005; 175(7):4265-73. PMC: 1378112. DOI: 10.4049/jimmunol.175.7.4265. View

Vodyanik M, Thomson J, Slukvin I . Leukosialin (CD43) defines hematopoietic progenitors in human embryonic stem cell differentiation cultures. Blood. 2006; 108(6):2095-105. PMC: 1895535. DOI: 10.1182/blood-2006-02-003327. View

Wang L, Menendez P, Cerdan C, Bhatia M . Hematopoietic development from human embryonic stem cell lines. Exp Hematol. 2005; 33(9):987-96. DOI: 10.1016/j.exphem.2005.06.002. View

Trounson A . The production and directed differentiation of human embryonic stem cells. Endocr Rev. 2006; 27(2):208-19. DOI: 10.1210/er.2005-0016. View

Goerdt S, Orfanos C . Other functions, other genes: alternative activation of antigen-presenting cells. Immunity. 1999; 10(2):137-42. DOI: 10.1016/s1074-7613(00)80014-x. View

Karlsson K, Cowley S, Martinez F, Shaw M, Minger S, James W . Homogeneous monocytes and macrophages from human embryonic stem cells following coculture-free differentiation in M-CSF and IL-3. Exp Hematol. 2008; 36(9):1167-75. PMC: 2635571. DOI: 10.1016/j.exphem.2008.04.009. View

Ihanus E, Uotila L, Toivanen A, Varis M, Gahmberg C . Red-cell ICAM-4 is a ligand for the monocyte/macrophage integrin CD11c/CD18: characterization of the binding sites on ICAM-4. Blood. 2006; 109(2):802-10. DOI: 10.1182/blood-2006-04-014878. View

Ingalls R, Golenbock D . CD11c/CD18, a transmembrane signaling receptor for lipopolysaccharide. J Exp Med. 1995; 181(4):1473-9. PMC: 2191975. DOI: 10.1084/jem.181.4.1473. View

10.

Zhan X, Dravid G, Ye Z, Hammond H, Shamblott M, Gearhart J . Functional antigen-presenting leucocytes derived from human embryonic stem cells in vitro. Lancet. 2004; 364(9429):163-71. DOI: 10.1016/S0140-6736(04)16629-4. View

11.

Ramprasad M, Terpstra V, Kondratenko N, Quehenberger O, Steinberg D . Cell surface expression of mouse macrosialin and human CD68 and their role as macrophage receptors for oxidized low density lipoprotein. Proc Natl Acad Sci U S A. 1996; 93(25):14833-8. PMC: 26222. DOI: 10.1073/pnas.93.25.14833. View

12.

Odorico J, Kaufman D, Thomson J . Multilineage differentiation from human embryonic stem cell lines. Stem Cells. 2001; 19(3):193-204. DOI: 10.1634/stemcells.19-3-193. View

13.

Doyle S, OConnell R, Miranda G, Vaidya S, Chow E, Liu P . Toll-like receptors induce a phagocytic gene program through p38. J Exp Med. 2003; 199(1):81-90. PMC: 1887723. DOI: 10.1084/jem.20031237. View

14.

Itskovitz-Eldor J, Schuldiner M, Karsenti D, Eden A, Yanuka O, Amit M . Differentiation of human embryonic stem cells into embryoid bodies compromising the three embryonic germ layers. Mol Med. 2000; 6(2):88-95. PMC: 1949933. View

15.

Chomarat P, Banchereau J, Davoust J, Palucka A . IL-6 switches the differentiation of monocytes from dendritic cells to macrophages. Nat Immunol. 2001; 1(6):510-4. DOI: 10.1038/82763. View

16.

Holness C, Simmons D . Molecular cloning of CD68, a human macrophage marker related to lysosomal glycoproteins. Blood. 1993; 81(6):1607-13. View

17.

Galic Z, Kitchen S, Kacena A, Subramanian A, Burke B, Cortado R . T lineage differentiation from human embryonic stem cells. Proc Natl Acad Sci U S A. 2006; 103(31):11742-7. PMC: 1544240. DOI: 10.1073/pnas.0604244103. View

18.

Peiser L, Mukhopadhyay S, Gordon S . Scavenger receptors in innate immunity. Curr Opin Immunol. 2002; 14(1):123-8. DOI: 10.1016/s0952-7915(01)00307-7. View

19.

Miranda M, Johnson D . Signal transduction pathways that contribute to myeloid differentiation. Leukemia. 2007; 21(7):1363-77. DOI: 10.1038/sj.leu.2404690. View

20.

Wang L, Li L, Shojaei F, Levac K, Cerdan C, Menendez P . Endothelial and hematopoietic cell fate of human embryonic stem cells originates from primitive endothelium with hemangioblastic properties. Immunity. 2004; 21(1):31-41. DOI: 10.1016/j.immuni.2004.06.006. View