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In vitro self-renewal division of hematopoietic stem cells.

Ema H, Takano H, Sudo K, Nakauchi H - J. Exp. Med. (2000)

Bottom Line: 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.This study provides evidence that one HSC regenerates at least one HSC in culture.

View Article: PubMed Central - PubMed

Affiliation: 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.

ABSTRACT
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.

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Cell division and colony formation of single CD34−KSL cells. A single-cell culture was performed in the presence of the indicated cytokines. The histogram shows the mean percentages of cells that underwent cell division at least once and of cells that gave rise to colonies during 2 wk of culture.
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Figure 1: Cell division and colony formation of single CD34−KSL cells. A single-cell culture was performed in the presence of the indicated cytokines. The histogram shows the mean percentages of cells that underwent cell division at least once and of cells that gave rise to colonies during 2 wk of culture.

Mentions: Direct effects of various cytokines on cell division of CD34−KSL cells were evaluated at the single-cell level in serum-free culture. Representative results for cell division and colony formation are graphically demonstrated in Fig. 1. SCF alone stimulated ∼25% of these cells to divide during 2 wk but did not further support them to form colonies consisting of >50 cells. Fewer than 10 or 5% of the cells underwent cell division on stimulation with TPO or IL-3 alone. No cell division was detected in culture with IL-6, IL-11, or G-CSF (data for IL-11 and G-CSF not shown).


In vitro self-renewal division of hematopoietic stem cells.

Ema H, Takano H, Sudo K, Nakauchi H - J. Exp. Med. (2000)

Cell division and colony formation of single CD34−KSL cells. A single-cell culture was performed in the presence of the indicated cytokines. The histogram shows the mean percentages of cells that underwent cell division at least once and of cells that gave rise to colonies during 2 wk of culture.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2193353&req=5

Figure 1: Cell division and colony formation of single CD34−KSL cells. A single-cell culture was performed in the presence of the indicated cytokines. The histogram shows the mean percentages of cells that underwent cell division at least once and of cells that gave rise to colonies during 2 wk of culture.
Mentions: Direct effects of various cytokines on cell division of CD34−KSL cells were evaluated at the single-cell level in serum-free culture. Representative results for cell division and colony formation are graphically demonstrated in Fig. 1. SCF alone stimulated ∼25% of these cells to divide during 2 wk but did not further support them to form colonies consisting of >50 cells. Fewer than 10 or 5% of the cells underwent cell division on stimulation with TPO or IL-3 alone. No cell division was detected in culture with IL-6, IL-11, or G-CSF (data for IL-11 and G-CSF not shown).

Bottom Line: 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.This study provides evidence that one HSC regenerates at least one HSC in culture.

View Article: PubMed Central - PubMed

Affiliation: 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.

ABSTRACT
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.

Show MeSH
Related in: MedlinePlus