Abstract
Hematopoietic stem cells (HSCs) can differentiate into several types of hematopoietic cells, such as erythrocytes, megakaryocytes, lymphocytes, neutrophils, or macrophages, and also undergo self-renewal to sustain hematopoiesis throughout an organism’s lifetime. HSCs emerge and expand during mouse embryogenesis. HSC regulation is governed by two types of activity: intrinsic activity programmed primarily by cell autonomous gene expression, and extrinsic factors, which originate from the so-called niche cells surrounding HSCs. Previously, we reported that endothelial niche cells regulate HSC generation at aorta-gonad-mesonephros region and placenta, and that hepatoblastic niche cells regulate HSC differentiation in mouse embryonic liver. In the course of those studies, we employed immunohistochemistry, flow cytometry, and the laser capture microdissection system to assess embryonic regulation of the mouse hematopoietic niche.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Dzierzak E, Medvinsky A, de Bruijn M (1998) Qualitative and quantitative aspects of hematopoietic cell development in the mammalian embryo. Immunol Today 19:228–236
McGrath KE, Palis J (2005) Hematopoiesis in the yolk sac: more than meets the eye. Exp Hematol 33:1021–1028
Ferkowicz MJ, Yoder MC (2005) Blood island formation: longstanding observations and modern interpretations. Exp Hematol 33:1041–1047
Sugiyama D, Tsuji K (2006) Definitive hematopoiesis from endothelial cells in the mouse embryo; a simple guide. Trends Cardiovasc Med 16:45–49
Dzierzak E, Speck NA (2008) Of lineage and legacy: the development of mammalian hematopoietic stem cells. Nat Immunol 9:129–136
Palis J, Robertson S, Kennedy M, Wall C, Keller G (1999) Development of erythroid and myeloid progenitors in the yolk sac and embryo proper of the mouse. Development 126:5073–5084
Medvinsky A, Dzierzak E (1996) Definitive hematopoiesis is autonomously initiated by the AGM region. Cell 86:897–906
Cumano A, Dieterlen-Lievre F, Godin I (1996) Lymphoid potential, probed before circulation in mouse, is restricted to caudal intraembryonic splanchnopleura. Cell 86:907–916
Yoder MC, Hiatt K, Dutt P, Mukherjee P, Bodine DM, Orlic D (1997) Characterization of definitive lymphohematopoietic stem cells in the day 9 murine yolk sac. Immunity 7:335–344
Matsuoka S, Tsuji K, Hisakawa H, Xu M, Ebihara Y, Ishii T, Sugiyama D, Manabe A, Tanaka R, Ikeda Y et al (2001) Generation of definitive hematopoietic stem cells from murine early yolk sac and paraaortic splanchnopleures by aorta-gonad-mesonephros region-derived stromal cells. Blood 98:6–12
Sugiyama D, Ogawa M, Nakao K, Osumi N, Nishikawa S, Nishikawa S, Arai K, Nakahata T, Tsuji K (2007) B cell potential can be obtained from pre-circulatory yolk sac, but with low frequency. Dev Biol 301:53–61
Samokhvalov IM, Samokhvalova NI, Nishikawa S (2007) Cell tracing shows the contribution of the yolk sac to adult haematopoiesis. Nature 446:1056–1061
Mikkola HK, Gekas C, Orkin SH, Dieterlen-Lievre F (2005) Placenta as a site for hematopoietic stem cell development. Exp Hematol 33:1048–1054
Zeigler BM, Sugiyama D, Chen M, Guo Y, Downs KM, Speck NA (2006) The allantois and chorion, when isolated before circulation or chorio-allantoic fusion, have hematopoietic potential. Development 133:4183–4192
Sasaki T, Mizuochi C, Horio Y, Nakao K, Akashi K, Sugiyama D (2010) Regulation of hematopoietic cell clusters in the placental niche through SCF/Kit signaling in embryonic mouse. Development 137:3941–3952
Johnson GR, Moore MA (1975) Role of stem cell migration in initiation of mouse foetal liver haemopoiesis. Nature 258:726–728
Houssaint E (1981) Differentiation of the mouse hepatic primordium. II. Extrinsic origin of the haemopoietic cell line. Cell Differ 10:243–252
Ema H, Nakauchi H (2000) Expansion of hematopoietic stem cells in the developing liver of a mouse embryo. Blood 95:2284–2288
Sugiyama D, Arai K, Tsuji K (2005) Definitive hematopoiesis from acetyl LDL incorporating endothelial cells in the mouse embryo. Stem Cells Dev 14:687–696
Zhang CC, Kaba M, Ge G, Xie K, Tong W, Hug C, Lodish HF (2006) Angiopoietin-like proteins stimulate ex vivo expansion of hematopoietic stem cells. Nat Med 12:240–245
Sugiyama D, Kulkeaw K, Mizuochi C, Horio Y, Okayama S (2011) Hepatoblasts comprise a niche for fetal liver erythropoiesis through cytokine production. Biochem Biophys Res Commun 410:301–306
Sugiyama D, Kulkeaw K, Mizuochi C (2013) TGF-beta-1 up-regulates extra-cellular matrix production in mouse hepatoblasts. Mech Dev 130(2–3):195–206
Mizuochi C, Fraser ST, Biasch K, Horio Y, Kikushige Y, Tani K, Akashi K, Tavian M, Sugiyama D (2012) Intra-aortic clusters undergo endothelial to hematopoietic phenotypic transition during early embryogenesis. PLoS One 7:e35763
Acknowledgments
We thank The Ministry of Education, Culture, Sports, Science and Technology; The Ministry of Health, Labor and Welfare; and The Japan Society for the Promotion of Science for grant support, and Dr. Elise Lamar for critical reading of the manuscript.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Sugiyama, D., Sasaki, T. (2013). Isolation of Embryonic Hematopoietic Niche Cells by Flow Cytometry and Laser Capture Microdissection. In: Turksen, K. (eds) Stem Cell Niche. Methods in Molecular Biology, vol 1035. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-508-8_6
Download citation
DOI: https://doi.org/10.1007/978-1-62703-508-8_6
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-507-1
Online ISBN: 978-1-62703-508-8
eBook Packages: Springer Protocols