The Journal of Experimental Medicine
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Published online 6 November 2000.
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© The Rockefeller University Press, 0022-1007/2000/11/1281/ $5.00
The Journal of Experimental Medicine, Volume 192, Number 9, November 6, 2000 1281-1288

Original Article

In Vitro Self-Renewal Division of Hematopoietic Stem Cells

Hideo Emaa, Hina Takanoa, Kazuhiro Sudoa, and Hiromitsu Nakauchia

a Department of Immunology, Institute of Basic Medical Sciences, University of Tsukuba and Core Research for Evolutional Science and Technology, Japan Science and Technology Corporation, Tsukuba 305-8575, Japan
Dept. of Immunology, Institute of Basic Medical Sciences, University of Tsukuba and CREST (JST), 1-1-1 Tennodai, Tsukuba 305-8575, Japan.81-298-53-696681-298-53-6964

Little is known about how hematopoietic stem cells (HSCs) self-renew. We studied the regeneration of HSCs in culture. Effects of various cytokines on cell division of CD34–/low c-Kit+Sca-1+ lineage marker–negative (CD34–KSL) bone marrow cells of the mouse were first evaluated in serum-free single cell culture. We then performed a competitive repopulation assay on divided cells to ask if such cell division involved self-renewal of HSCs.

In the presence of stem cell factor (SCF), thrombopoietin (TPO) induced a first cell division of CD34–KSL cells more efficiently than did interleukin (IL)-3 or IL-6. Multilineage repopulating cells were detected in a significant proportion of cells derived from single cells in culture with TPO and SCF, although this culture condition led to a substantial decrease in HSC number. These regenerated repopulating cells could be further transplanted into secondary recipients. When paired daughter cells were separately studied, one of a pair gave rise to repopulating cells with self-renewal potential, suggesting asymmetric self-renewal division. This study provides evidence that one HSC regenerates at least one HSC in culture.

Key Words: mouse • bone marrow transplantation • hematopoiesis • self-renewal • cell division

Abbreviations used in this paper: HSCs, hematopoietic stem cells; SCF, stem cell factor; TPO, thrombopoietin.

© 2000 The Rockefeller University Press


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