In vitro assays for primitive hematopoietic cells [letter; comment]

Ploemacher, R. E.; Loo, JC van der; Sluijs, JP Van der

doi:10.1182/blood.v79.3.834.834

Cited by 5 publications

(2 citation statements)

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“…We believe that this results from our use of more stringent criteria in the CAFC assay. As has been pointed out by Ploemacher et al (25), the CAFC assay will only result in reproducible and accurate stem cell measurements if (phase contrast-negative) colonies growing beneath stromal cells are counted. Phase contrast-positive cells accumulating on top of the stroma reflect the mature progeny of primitive cells, and their presence does not indicate a proper cobblestone area.…”

Section: Mapping Of a Locus Which Regulates The Population Size Of Primitive Stem Cell Compartmentsmentioning

confidence: 99%

Intrinsic and Extrinsic Control of Hemopoietic Stem Cell Numbers: Mapping of a Stem Cell Gene

Haan

Zant

1997

The Journal of Experimental Medicine

127

101

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We evaluated in vivo interactions between extrinsic (growth factor induced) and intrinsic (genetically determined) effectors of mouse primitive hemopoietic stem cell proliferation and numbers. Accordingly, stem cell frequency and cell cycle kinetics were assessed in eight strains of inbred mice using the cobblestone area–forming cell (CAFC) assay. A strong inverse correlation was observed between mouse lifespan and the number of autonomously cycling progenitors (CAFC day 7) in the femur. The population size of primitive stem cells (CAFC day 35) varied widely (up to sevenfold) among strains, unlike total CAFC day 7 numbers (cycling and quiescent), which were similar. Administration of the early acting cytokine flt-3 ligand to these strains resulted in activation of quiescent primitive stem cells exclusively in strains with high endogenous stem cell numbers (DBA and AKR), but was unrelated to strain-specific progenitor cell cycling. To map loci affecting stem cell frequency, we quantified stem cells in BXD recombinant inbred mice (offspring of C57BL/6 and DBA/2). The resulting strain distribution pattern showed high concordance with a marker that mapped to chromosome 18 (19 cM). Linkage with this genomic interval was associated with a likelihood of odds score of 3.3, surpassing the level required for significance. Interestingly, this segment, containing the EGR-1 gene, shows synteny with human chromosome 5q, a region strongly associated with various hematological malignancies. Our findings indicate that a gene mapping to this region is mutated in either C57BL/6 or DBA/2 (and possibly AKR) mice. These studies in apparently healthy mice may facilitate the identification of a gene implicated in human 5q-syndromes.

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Section: Mapping Of a Locus Which Regulates The Population Size Of Primitive Stem Cell Compartmentsmentioning

confidence: 99%

Intrinsic and Extrinsic Control of Hemopoietic Stem Cell Numbers: Mapping of a Stem Cell Gene

Haan

Zant

1997

The Journal of Experimental Medicine

127

101

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“…At the end of the experiment, the number of positively or negatively regenerated recipients is scored. The results are analyzed assuming that (1) only self-renewing cells regenerate the recipients, (2) one rare and randomly dispersed donor cell with self-renewing ability is enough to regenerate the recipient [ 4 ], and (3) the probability of regeneration follows a single-hit model of the Poisson distribution [ 5 ]. In addition, there are different in vitro serial dilution assays but since they may not recapitulate the entire tissue hierarchy, caution needs to be used when drawing conclusions about self-renewal from them.…”

Section: Understanding Self-renewal Is Critical To Study Tissue Rementioning

confidence: 99%

The Aryl Hydrocarbon Receptor Relays Metabolic Signals to Promote Cellular Regeneration

Casado

2016

Stem Cells International

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While sensing the cell environment, the aryl hydrocarbon receptor (AHR) interacts with different pathways involved in cellular homeostasis. This review summarizes evidence suggesting that cellular regeneration in the context of aging and diseases can be modulated by AHR signaling on stem cells. New insights connect orphaned observations into AHR interactions with critical signaling pathways such as WNT to propose a role of this ligand-activated transcription factor in the modulation of cellular regeneration by altering pathways that nurture cellular expansion such as changes in the metabolic efficiency rather than by directly altering cell cycling, proliferation, or cell death. Targeting the AHR to promote regeneration might prove to be a useful strategy to avoid unbalanced disruptions of homeostasis that may promote disease and also provide biological rationale for potential regenerative medicine approaches.

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Non‐hematopoietic human bone marrow contains long‐lasting, pluripotential mesenchymal stem cells

Suvà

Garavaglia

Ménétrey

et al. 2003

Journal Cellular Physiology

115

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Mesenchymal stem cells (MSC) are considered as potential agents for reconstructive and gene-targeting therapies since they differentiate into various cell-lineages, exhibit an extended survival once injected into a host, and can easily be transfected with engineered DNA. MSC are essentially isolated from hematopoietic bone marrow (BM), a process that is rather invasive and may raise ethical concerns. In an attempt to find an alternative source, we evaluated whether non-hematopoietic (nh)BM recovered from femoral heads of patients undergoing hip arthroplasty contained MSC. Ex vivo, 99% of nhBM cells were CD45(+) leukocytes. After culture, leukocytes were replaced by a homogeneous layer of adherent CD45(-) CD14(-) CD34(-) CD11b(-) CD90(+) HLA-ABC(+) cells. Culture doubling time (mean = 4 days, range 1.6-6.7 days) was not correlated with patient age (27-81 years, n = 16). Amplified cultures supported long-term hematopoiesis, and could be differentiated in vitro into adipocytes and chondrocytes. Moreover, a small fraction of nhBM cells spontaneously expressed MyoD1 and formed myotubes, suggesting that myogenic differentiation also occurred. nhBM contained clonogenic cells whose frequency (1/13,000), doubling time (2.1 days), and maximal amplification (up to 10(6)-fold) were not age-related. All 14 clones analyzed (from five patients, ages 27-78 years) differentiated into at least one mesenchymal lineage, and 66% were bipotential (n = 8/12), or tripotential (n = 2/3). In conclusion, nhBM contains pluripotential mesenchymal progenitors which are similar to hematopoietic BM-derived MSC, and whose biological functions are not altered by aging. Furthermore, if MSC-based therapies hold their promises, nhBM may become the source of choice for responding to the increasing demand for MSC.

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In vitro assays for primitive hematopoietic cells [letter; comment]

Cited by 5 publications

References 0 publications

Intrinsic and Extrinsic Control of Hemopoietic Stem Cell Numbers: Mapping of a Stem Cell Gene

Intrinsic and Extrinsic Control of Hemopoietic Stem Cell Numbers: Mapping of a Stem Cell Gene

The Aryl Hydrocarbon Receptor Relays Metabolic Signals to Promote Cellular Regeneration

Non‐hematopoietic human bone marrow contains long‐lasting, pluripotential mesenchymal stem cells

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