Expansion on Stromal Cells Preserves the Undifferentiated State of Human Hematopoietic Stem Cells Despite Compromised Reconstitution Ability

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2013 Jan 24

PMID 23342037

Citations 15

Authors

Mattias Magnusson

Maria I Sierra

Rajkumar Sasidharan

Sacha L Prashad

Melissa Romero

Pamela Saarikoski

Ben Van Handel

Andy Huang

Xinmin Li

Hanna K A Mikkola

Affiliations

Soon will be listed here.

Abstract

Lack of HLA-matched hematopoietic stem cells (HSC) limits the number of patients with life-threatening blood disorders that can be treated by HSC transplantation. So far, insufficient understanding of the regulatory mechanisms governing human HSC has precluded the development of effective protocols for culturing HSC for therapeutic use and molecular studies. We defined a culture system using OP9M2 mesenchymal stem cell (MSC) stroma that protects human hematopoietic stem/progenitor cells (HSPC) from differentiation and apoptosis. In addition, it facilitates a dramatic expansion of multipotent progenitors that retain the immunophenotype (CD34+CD38-CD90+) characteristic of human HSPC and proliferative potential over several weeks in culture. In contrast, transplantable HSC could be maintained, but not significantly expanded, during 2-week culture. Temporal analysis of the transcriptome of the ex vivo expanded CD34+CD38-CD90+ cells documented remarkable stability of most transcriptional regulators known to govern the undifferentiated HSC state. Nevertheless, it revealed dynamic fluctuations in transcriptional programs that associate with HSC behavior and may compromise HSC function, such as dysregulation of PBX1 regulated genetic networks. This culture system serves now as a platform for modeling human multilineage hematopoietic stem/progenitor cell hierarchy and studying the complex regulation of HSC identity and function required for successful ex vivo expansion of transplantable HSC.

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References

Le Blanc K, Ringden O . Immunomodulation by mesenchymal stem cells and clinical experience. J Intern Med. 2007; 262(5):509-25. DOI: 10.1111/j.1365-2796.2007.01844.x. View

Jude C, Gaudet J, Speck N, Ernst P . Leukemia and hematopoietic stem cells: balancing proliferation and quiescence. Cell Cycle. 2008; 7(5):586-91. PMC: 2892629. DOI: 10.4161/cc.7.5.5549. View

Collins L, Dorshkind K . A stromal cell line from myeloid long-term bone marrow cultures can support myelopoiesis and B lymphopoiesis. J Immunol. 1987; 138(4):1082-7. View

Pevny L, Simon M, Robertson E, Klein W, Tsai S, DAgati V . Erythroid differentiation in chimaeric mice blocked by a targeted mutation in the gene for transcription factor GATA-1. Nature. 1991; 349(6306):257-60. DOI: 10.1038/349257a0. View

Miller C, Eaves C . Expansion in vitro of adult murine hematopoietic stem cells with transplantable lympho-myeloid reconstituting ability. Proc Natl Acad Sci U S A. 1998; 94(25):13648-53. PMC: 28360. DOI: 10.1073/pnas.94.25.13648. View

Amsellem S, Pflumio F, Bardinet D, Izac B, Charneau P, Romeo P . Ex vivo expansion of human hematopoietic stem cells by direct delivery of the HOXB4 homeoprotein. Nat Med. 2003; 9(11):1423-7. DOI: 10.1038/nm953. View

Krosl J, Beslu N, Mayotte N, Humphries R, Sauvageau G . The competitive nature of HOXB4-transduced HSC is limited by PBX1: the generation of ultra-competitive stem cells retaining full differentiation potential. Immunity. 2003; 18(4):561-71. DOI: 10.1016/s1074-7613(03)00090-6. View

Buza-Vidas N, Duarte S, Luc S, Bouriez-Jones T, Woll P, Jacobsen S . GATA3 is redundant for maintenance and self-renewal of hematopoietic stem cells. Blood. 2011; 118(5):1291-3. DOI: 10.1182/blood-2011-02-338046. View

Dorrell C, Gan O, Pereira D, Hawley R, Dick J . Expansion of human cord blood CD34(+)CD38(-) cells in ex vivo culture during retroviral transduction without a corresponding increase in SCID repopulating cell (SRC) frequency: dissociation of SRC phenotype and function. Blood. 1999; 95(1):102-10. View

10.

Cai M, Langer E, Gill J, Satpathy A, Albring J, Kc W . Dual actions of Meis1 inhibit erythroid progenitor development and sustain general hematopoietic cell proliferation. Blood. 2012; 120(2):335-46. PMC: 3628121. DOI: 10.1182/blood-2012-01-403139. View

11.

Calvi L, Adams G, Weibrecht K, Weber J, Olson D, Knight M . Osteoblastic cells regulate the haematopoietic stem cell niche. Nature. 2003; 425(6960):841-6. DOI: 10.1038/nature02040. View

12.

Danet G, Pan Y, Luongo J, Bonnet D, Simon M . Expansion of human SCID-repopulating cells under hypoxic conditions. J Clin Invest. 2003; 112(1):126-35. PMC: 162287. DOI: 10.1172/JCI17669. View

13.

Challen G, Sun D, Jeong M, Luo M, Jelinek J, Berg J . Dnmt3a is essential for hematopoietic stem cell differentiation. Nat Genet. 2011; 44(1):23-31. PMC: 3637952. DOI: 10.1038/ng.1009. View

14.

Zhang C, Kaba M, Ge G, Xie K, Tong W, Hug C . Angiopoietin-like proteins stimulate ex vivo expansion of hematopoietic stem cells. Nat Med. 2006; 12(2):240-5. PMC: 2771412. DOI: 10.1038/nm1342. View

15.

Nakauchi H, Sudo K, Ema H . Quantitative assessment of the stem cell self-renewal capacity. Ann N Y Acad Sci. 2001; 938:18-24; discussion 24-5. DOI: 10.1111/j.1749-6632.2001.tb03570.x. View

16.

Sudo T, Ohno N, Akiyama N, Tamakoshi M, Yoshida H, Nishikawa S . Expression and function of the interleukin 7 receptor in murine lymphocytes. Proc Natl Acad Sci U S A. 1993; 90(19):9125-9. PMC: 47514. DOI: 10.1073/pnas.90.19.9125. View

17.

Oostendorp R, Harvey K, Kusadasi N, de Bruijn M, Saris C, Ploemacher R . Stromal cell lines from mouse aorta-gonads-mesonephros subregions are potent supporters of hematopoietic stem cell activity. Blood. 2002; 99(4):1183-9. DOI: 10.1182/blood.v99.4.1183. View

18.

Wettenhall J, Simpson K, Satterley K, Smyth G . affylmGUI: a graphical user interface for linear modeling of single channel microarray data. Bioinformatics. 2006; 22(7):897-9. DOI: 10.1093/bioinformatics/btl025. View

19.

Shimakura Y, Kawada H, Ando K, Sato T, Nakamura Y, Tsuji T . Murine stromal cell line HESS-5 maintains reconstituting ability of Ex vivo-generated hematopoietic stem cells from human bone marrow and cytokine-mobilized peripheral blood. Stem Cells. 2000; 18(3):183-9. DOI: 10.1634/stemcells.18-3-183. View

20.

Weissman I . Stem cells: units of development, units of regeneration, and units in evolution. Cell. 2000; 100(1):157-68. DOI: 10.1016/s0092-8674(00)81692-x. View