Effects of developmental age, brain region, and time in culture on long-term proliferation and multipotency of neural stem cell populations

The ability to determine the functional capacity of putative human hematopoietic stem cell (HSC) populations requires in vivo assays in which long-term multilineage differentiation can be assessed. We hypothesized that if human fetal bone was transplanted adjacent to a fetal thymus fragment in severe combined immunodeficient (SCID) mice, a conjoint organ might form in which HSC in the human bone marrow (BM) would mimic human multilineage differentiation into progenitor cells, B cells, and myeloid cells; undergo self-renewal; and migrate to and differentiate into T cells within the thymic microenvironment. To test this possibility, SCID mice were transplanted subcutaneously with HLA class I mismatched fetal bone, thymus, and spleen fragments (SCID-hu BTS). We found that the BM of SCID-hu BTS grafts maintained B cells, myeloid cells, CD34+ cells for at least 36 weeks posttransplant. Assayable hematopoietic progenitors colony-forming units-granulocyte- macrophage were present in 100% (66/66) of grafts over a period of 28 weeks. Cells with a HSC phenotype (CD34+Thy-1+Lin-) were maintained for 20 weeks in SCID-hu BTS grafts. These CD34+Thy-1+Lin- cells had potent secondary multilineage reconstituting potential when isolated and injected into a secondary HLA mismatched SCID-hu bone assay and analyzed 8 weeks later. In addition, early progenitors within the BM of SCID-hu BTS grafts were capable of migrating to the human thymus and undergoing differentiation through immature CD4+CD8+ double-positive T cells and produce mature T cells with a CD4+CD8- or CD8+CD4- phenotype that could be detected for at least 36 weeks. Phenotypically defined human fetal liver (FL) and umbilical cord blood (UCB) hematopoietic stem cell populations were injected into irradiated SCID-hu BTS grafts to assess their multilineage repopulating capacity and to assess the ability of the BTS system to provide an environment where multiple lineages might differentiate from a common stem cell pool. Injection of irradiated grafts with FL HSC or UCB HSC cells resulted in donor- derived B cells, myeloid cells, immature and mature T cells, and CD34+ cells in individual grafts when analyzed 8 weeks postreconstitution, further showing the multipotential nature of these stem cell populations. In addition, a strong correlation was observed between maintenance of host graft-derived CD8+ cells and failure of donor stem cell engraftment.(ABSTRACT TRUNCATED AT 400 WORDS)

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Retinoid Machinery in Distinct Neural Stem Cell Populations with Different Retinoid Responsiveness

Stem Cells and Development ◽

10.1089/scd.2012.0422 ◽

2013 ◽

Vol 22 (20) ◽

pp. 2777-2793 ◽

Cited By ~ 9

Author(s):

Barbara Orsolits ◽

Adrienn Borsy ◽

Emília Madarász ◽

Zsófia Mészáros ◽

Tímea Kőhidi ◽

...

Keyword(s):

Stem Cell ◽

Neural Stem Cell ◽

Cell Populations ◽

Stem Cell Populations

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Long Term, Low Dose Genistein decreases Stem Cell Populations and Sensitizes Inflammatory Breast Cancer Cell Lines to Radiation

Cancer Studies and Molecular Medicine - Open Journal ◽

10.17140/csmmoj-2-107 ◽

2015 ◽

Vol 2 (1) ◽

pp. 60-65 ◽

Cited By ~ 2

Author(s):

Jennifer Sims-Mourtada ◽

◽

Lynn M. Opdenaker ◽

Joshua Davis ◽

Changqing Wu ◽

...

Keyword(s):

Breast Cancer ◽

Stem Cell ◽

Breast Cancer Cell ◽

Inflammatory Breast Cancer ◽

Low Dose ◽

Breast Cancer Cell Lines ◽

Cell Populations ◽

Inflammatory Breast Cancer Cell ◽

Stem Cell Populations

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Stem Cells in Paediatric Leukaemia Are Resistant to Parthenolide.

Blood ◽

10.1182/blood.v114.22.1429.1429 ◽

2009 ◽

Vol 114 (22) ◽

pp. 1429-1429

Author(s):

Charlotte Victoria Cox ◽

Paraskevi Diamanti ◽

Allison Blair

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Functional Capacity ◽

Cell Populations ◽

Minimal Effect ◽

Childhood All ◽

Stem Cell Populations

Abstract Abstract 1429 Poster Board I-452 The concept of cancer stem cells as developmentally early cells that are capable of continued growth and expansion in haematopoietic malignancies and solid tumours has been substantiated in recent years. Consequently these cells may be responsible for disease maintenance and relapse. Acute lymphoblastic leukaemia (ALL) is the most common paediatric cancer with survival rates around 80-85%. However, a significant proportion of patients relapse, often with disease that is highly refractory to further therapeutic intervention. Leukaemia stem cells have been described in childhood ALL that can proliferate to initiate and sustain the disease in vivo. In addition these leukaemia stem cells have also been shown to be refractory to commonly used clinical agents. Therefore it is important to investigate ALL stem cells to understand their biological properties and to identify the most appropriate agents that are capable of eradicating these cells. The sesquiterpene lactone Parthenolide (PTL) has been shown to induce apoptosis in malignant cells by inducing oxidative stress and inhibiting NF-κB activity. Importantly PTL has been shown to be effective against stem cell populations in acute myeloid leukaemia and in chronic lymphocytic leukaemia with minimal effect on normal haemopoietic cells. In this study we have attempted to assess the effects of PTL on stem cell populations in paediatric ALL. Primary cells from 20 childhood ALL cases from mixed prognostic subgroups were used in this investigation. Cells from B-ALL cases were sorted on the basis of expression of CD34/CD19, while CD34/CD7 antigens were used to sort cells from T-ALL cases. Sorted and unsorted populations were co-cultured with and without PTL at 7.5μM and 10μM for 18-24 hours. Subsequently cell viability and apoptosis were determined by flow cytometry using Annexin V and PI staining. Antibodies against phosphorylated IKKα and IKKβ were used to assess NF-κB activity in treated and untreated cells. The functional ability of the treated cells was assessed in some cases using long-term in vitro and in vivo assays. Both concentrations of PTL resulted in a significant reduction in viability in unsorted ALL cells (28±4% and 23±5% respectively). Similar results were observed with CD34+/CD19+, CD34+/CD7+ and CD34- subfractions, with viability reduced to 14-39%. In contrast the phenotypically primitive CD34+/CD19- (85±11% viable) and CD34+/CD7- (83±5% viable) populations were significantly more resistant to 10μM PTL than unsorted cells and other sorted populations (P≤0.002). FISH analyses were performed at the end of the time-course and confirmed that leukaemia cells were surviving PTL treatment. It was not possible to detect phosphorylated IKKα/β in the CD34+/CD19- and CD34+/CD7- populations, in cases examined to date, suggesting NF-kB may not be active in these subpopulations. Of note PTL treatment seemed to have minimal effect on the long-term proliferative ability of ALL cells. There were no significant differences in the absolute cell numbers generated in cultures of PTL treated CD34+/CD19- or CD34+/CD7- cells compared to untreated cells at all time points assayed up to the end of culture at week 6 (P≥0.23). Interestingly, similar results were observed with the unsorted cells and all other sorted populations. From week 3 of culture there was no difference in the absolute cell counts when growth from treated and untreated cells was compared (P>0.47), albeit they proliferated to a much lesser extent than the phenotypically primitive populations. In addition PTL treated cells were capable of engrafting NOD/SCID mice. The levels of leukaemia engraftment obtained using PTL treated unsorted (0.2-5% CD45+), CD34+/CD19- (2-10% CD45+) and CD34+/CD7- (1.5-9% CD45+) populations were similar to their respective untreated controls. These data demonstrate that while PTL showed promising effects on the bulk leukaemia cells, the effects on CD34+/CD19- B-ALL cells and CD34+/CD7- T-ALL cells were insignificant. This may be due in part to lack of NF-kB activity in leukaemia stem cells. However, the functional capacity of every ALL population evaluated in vitro was not significantly impaired by the short course of PTL treatment. These findings further highlight the importance of evaluating new therapeutic agents on leukaemia stem cell populations in addition to the bulk leukaemia and the significance of investigating the functional capacity of drug treated cells. Disclosures No relevant conflicts of interest to declare.

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Designing signaling environments to steer transcriptional diversity in neural progenitor cell populations

10.1101/2019.12.30.890087 ◽

2019 ◽

Author(s):

Jong H. Park ◽

Tiffany Tsou ◽

Paul Rivaud ◽

Matt Thomson ◽

Sisi Chen

Keyword(s):

Cell Culture ◽

Population Structure ◽

Stem Cell ◽

Single Cell ◽

Neural Stem Cell ◽

Cell Populations ◽

List Type ◽

Developmental Potential ◽

Stem Cell Populations

AbstractStem cell populations within developing embryos are diverse, composed of many different sub-populations of cells with varying developmental potential. The structure of stem cell populations in cell culture remains poorly understood and presents a barrier to differentiating stem cells for therapeutic applications. In this paper we develop a framework for controlling the architecture of stem cell populations in cell culture using high-throughput single cell mRNA-seq and computational analysis. We find that the transcriptional diversity of neural stem cell populations collapses in cell culture. Cell populations are depleted of committed neuron progenitor cells and become dominated by a single pre-astrocytic cell population. By analyzing the response of neural stem cell populations to forty distinct signaling conditions, we demonstrate that signaling environments can restructure cell populations by modulating the relative abundance of pre-astrocyte and pre-neuron subpopulations according to a simple linear code. One specific combination of BMP4, EGF, and FGF2 ligands switches the default population balance such that 70% of cells correspond to the committed neurons. Our work demonstrates that single-cell RNA-seq can be applied to modulate the diversity of in vitro stem cell populations providing a new strategy for population-level stem cell control.HighlightsNatural progenitor diversity in the brain collapses during in vitro culture to a single progenitor typeLoss of progenitor diversity alters fate potential of cells during differentiationLarge scale single-cell signaling screen identifies signals that reshape population structure towards neuronal cell typesSignals regulate population structure according to a simple log-linear model

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