Self-renewal of hemopoietic stem cells during mixed colony formation in vitro.

Humphries, R. Keith; Eaves, AC; Eaves, Connie J.

doi:10.1073/pnas.78.6.3629

Cited by 136 publications

(90 citation statements)

References 21 publications

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“…These findings are consistent with a more rapid loss in vitro of the HSC subtypes that retain extensive self-renewal activity in vivo 10 and a stochastic distribution of self-renewal events among individual multipotent cells proliferating in vitro. 51 At the same time, they appear inconsistent with the reported symmetry of the first 2 divisions of CD34 Ϫ KSL cells in vitro 52 and of the clonally amplified HSCs seen after 10 days in vitro. 10 Clearly, additional experiments will be required to determine more precisely when and how symmetry is established when HSCs are stimulated under different conditions in vitro.…”

Section: Discussioncontrasting

confidence: 53%

Steel factor coordinately regulates the molecular signature and biologic function of hematopoietic stem cells

Kent

Dykstra

Cheyne

et al. 2008

Blood

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Hematopoietic stem cells (HSCs) regenerated in vivo display sustained differences in their self-renewal and differentiation activities. Variations in Steel factor (SF) signaling are known to affect these functions in vitro, but the cellular and molecular mechanisms involved are not understood. To address these issues, we evaluated highly purified HSCs maintained in single-cell serum-free cultures containing 20 ng/mL IL-11 plus 1, 10, or 300 ng/mL SF. Under all conditions, more than 99% of the cells traversed a first cell cycle with similar kinetics. After 8 hours in the 10 or 300 ng/mL SF conditions, the frequency of HSCs remained unchanged. However, in the next 8 hours (ie, 6 hours before any cell divided), HSC integrity was sustained only in the 300 ng/mL SF cultures. The cells in these cultures also contained significantly higher levels of Bmi1, Lnk, and Ezh2 transcripts but not of several other regulators. Assessment of 21 first division progeny pairs further showed that only those generated in 300 ng/mL SF cultures contained HSCs and pairs of progeny with similar differentiation programs were not observed. Thus, SF signaling intensity can directly and coordinately alter the transcription factor profile and long-term repopulating ability of quiescent HSCs before their first division. (Blood. 2008;112:560-567) IntroductionThe hematopoietic system of the adult mouse is responsible for the daily production of billions of differentiated blood cells of various types. Because most of these cells have a limited lifespan and proliferative ability, they must be continuously generated from a population of more primitive cells that collectively have life-long self-sustaining ability. This function is restricted to a tiny subset of multipotent cells generally referred to as hematopoietic stem cells (HSCs). Historically, HSCs have been both defined and quantified retrospectively by their ability to generate clones containing both lymphoid and myeloid blood cells for at least 4 months when transplanted into irradiated recipients at limiting dilutions. 1 Analyses of such mice have also allowed the long-term differentiation activity of individual HSCs to be characterized. 2,3 Methods have now been developed for isolating purified populations in which 20% to 60% of the cells display this durability of reconstituting activity, indicating that intravenously injected HSCs can have very high seeding efficiencies in irradiated mice. [4][5][6][7] The ability to isolate such highly purified HSC populations has also permitted a more direct and complete analysis of their in vivo differentiation activity in single-cell transplant experiments. 6,[8][9][10][11] More recently, serial transplants of such clonally repopulated mice have been performed. Together, these experiments have revealed that HSCs possess one of 4 distinct differentiation programs that are propagated over many generations in vivo. 10,12 In addition, these studies have shown that extensive self-renewal ability in vivo is strongly associated with the display of either a...

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Section: Discussioncontrasting

confidence: 53%

Steel factor coordinately regulates the molecular signature and biologic function of hematopoietic stem cells

Kent

Dykstra

Cheyne

et al. 2008

Blood

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“…Stochastic models previously have been invoked to explain commitment to terminal differentiation in induced mouse erythroleukemia cells (11,12) and the process of commitment and self-renewal of murine pluripotent stem cells in vivo or in vitro (13,14). The observations and the analyses described in this paper strongly suggest that the expression of Hb F in the adult erythroid cultures is also controlled by a cellular process that is stochastic.…”

Section: Discussionmentioning

confidence: 48%

Stochastic expression of fetal hemoglobin in adult erythroid cells.

Stamatoyannopoulos¹,

Kurnit²,

Papayannopoulou³

1981

Proc. Natl. Acad. Sci. U.S.A.

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The expression of fetal hemoglobin, Hb F, in the adult cells is cellularly restricted both in vivo and in culture. Because, in cultures of erythroid progenitors, subclones that express or fail to express Hb F are derived from the same erythroid stem cell, a mechanism must exist whereby Hb F expression segregates in the progeny of erythroid progenitors during their differentiation. We present mathematical analyses of experimental data which suggest that expression of Hb F in human adult erythroid cells occurs on a stochastic basis. We quantified Hb F expression among the subclones of erythroid bursts (clones) in vitro by labeling subclones with fluorescent anti-Hb F antibodies. The observed data were compared with predictions from a stochastic model with the assumption that the expression of Hb F in a subclone occurs with a probability P equal to the frequency of Hb F-expressing subclones in an experiment. There was good fit between the observed and predicted data with respect to: (i) the relative frequencies of monomorphic F+, monomorphic F-, and bimorphic F+/F- bursts, respectively; (ii) the size distributions among F+, F- and F+/F- bursts; and (iii) the proportions of subclones expressing Hb F within bimorphic F+/F- bursts. Given the hypothesis that erythroid progenitors which have an active Hb F program are less differentiated than cells which do not proceed to express Hb F, the stochastic event indicated by our analyses may be the probability that adult erythroid progenitor cells undergo terminal differentiation at an earlier stage than usual.

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“…Spleen cells from animals transplanted with HOXB4-neo-or neo-transduced cells were plated 3x10 s cells/dish, or 3x 106 cells/dish, respectively. Colonies were scored on day 12 to day 14 of incubation as derived from CFU-GM, BFU-E, or CFU-GEMM according to standard criteria (Humphries et al 1981). …”

Section: In V/tro Clonogenic Progenitor Assaysmentioning

confidence: 99%

Overexpression of HOXB4 in hematopoietic cells causes the selective expansion of more primitive populations in vitro and in vivo.

Sauvageau

Þorsteinsdóttir²,

Eaves³

et al. 1995

Genes Dev.

Self Cite

548

467

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Hox genes were first recognized for their role in embryonic development and may also play important lineage-specific functions in a variety of somatic tissues including the hematopoietic system. We have recently shown that certain members of the Hox A and B clusters, such as HOXB3 and HOXB4, are preferentially expressed in subpopulations of human bone marrow that are highly enriched for the most primitive hematopoietic cell types. To assess the role these genes may play in regulating the proliferation and/or differentiation of such cells, we engineered the overexpression of HOXB4 in murine bone marrow cells by retroviral gene transfer and analyzed subsequent effects on the behavior of various hematopoietic stem and progenitor cell populations both in vitro and in vivo. Serial transplantation studies revealed a greatly enhanced ability of HOXB4.transduced bone marrow cells to regenerate the most primitive hematopoietic stem cell compartment resulting in 50-fold higher numbers of transplantable totipotent hematopoietic stem cells in primary and secondary recipients, compared with serially passaged neo-infected control cells. This heightened expansion in vivo of HOXB4.transduced hematopoietic stem cells was not accompanied by identifiable anomalies in the peripheral blood of these mice. Enhanced proliferation in vitro of day-12 CFU-S and clonogenic progenitors was also documented. These results indicate HOXB4 to be an important regulator of very early but not late hematopoietic cell proliferation and suggest a new approach to the controlled amplification of genetically modified hematopoietic stem cell populations.[Key Words: Homeo box genes~ bone marrow transplantation~ retroviral gene transfer]

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Self-renewal of hemopoietic stem cells during mixed colony formation in vitro.

Cited by 136 publications

References 21 publications

Steel factor coordinately regulates the molecular signature and biologic function of hematopoietic stem cells

Steel factor coordinately regulates the molecular signature and biologic function of hematopoietic stem cells

Stochastic expression of fetal hemoglobin in adult erythroid cells.

Overexpression of HOXB4 in hematopoietic cells causes the selective expansion of more primitive populations in vitro and in vivo.

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