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Developmental changes in the in vitro activated regenerative activity of primitive mammary epithelial cells

ABSTRACT: Many normal adult tissues contain rare stem cells with extensive self-maintaining regenerative potential. During development, the stem cells of the hematopoietic and neural systems undergo intrinsically specified changes in their self-renewal potential. In the mouse, mammary stem cells with transplantable regenerative activity are first detectable a few days before birth. They share some phenotypic properties with their adult counterparts but are enriched in a subpopulation that displays a distinct gene expression profile. Here we show that fetal mammary epithelial cells have a greater direct and inducible growth potential than their adult counterparts. The latter feature is revealed in a novel culture system that enables large numbers of mammary stem cells with serially transplantable activity as well as in vitro clonogenic progenitors to be produced within 7 days from single fetal or adult input cells. We further show that these responses are highly dependent on novel factors produced by fibroblasts. These findings provide new avenues for elucidating mechanisms that regulate normal mammary epithelial stem cell properties at the single-cell level, how these change during development, and how their perturbation may contribute to transformation. We used microarrays to compare the transcriptome of E18.5 fetal and adult MRU-enriched mammary cells.

ORGANISM(S): Mus musculus

PROVIDER: GSE46357 | GEO | 2013/08/13

SECONDARY ACCESSION(S): PRJNA198806

REPOSITORIES: GEO

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Developmental changes in the in vitro activated regenerative activity of primitive mammary epithelial cells

Project description:Many normal adult tissues contain rare stem cells with extensive self-maintaining regenerative potential. During development, the stem cells of the hematopoietic and neural systems undergo intrinsically specified changes in their self-renewal potential. In the mouse, mammary stem cells with transplantable regenerative activity are first detectable a few days before birth. They share some phenotypic properties with their adult counterparts but are enriched in a subpopulation that displays a distinct gene expression profile. Here we show that fetal mammary epithelial cells have a greater direct and inducible growth potential than their adult counterparts. The latter feature is revealed in a novel culture system that enables large numbers of mammary stem cells with serially transplantable activity as well as in vitro clonogenic progenitors to be produced within 7 days from single fetal or adult input cells. We further show that these responses are highly dependent on novel factors produced by fibroblasts. These findings provide new avenues for elucidating mechanisms that regulate normal mammary epithelial stem cell properties at the single-cell level, how these change during development, and how their perturbation may contribute to transformation. We used microarrays to compare the transcriptome of E18.5 fetal and adult MRU-enriched mammary cells. Three biological replicates each of CD31-CD45-Ter119-BP-1-EpCAM+CD49f+ adult basal cells and CD31-CD45-Ter119-EpCAM++CD49f+ fetal cells were sorted. RNA was extracted and hybridized to the Agilent One-Color Gene Expression Arrays .

2013-08-13 | E-GEOD-46357 | biostudies-arrayexpress

Expression profiling of mammary stem cell enriched fractions and other fractions from fetal and adult mouse.

2012-02-08 | GSE27027 | GEO

Expression profiling of mammary stem cell enriched fractions and other fractions from fetal and adult mouse.

Project description:Examination of genome-wide, gene expression level differences among adult and fetal mammary sub-populations including fetal CD24high cells and adult CD49fhighCD24medium cells which harbor enriched mammary stem cell activity. A genome wide 12-plex expression array using linear-amplified (Eberwine) mRNA from dissociated and FACS sorted fetal and adult mammary gland populations. Samples represent independent pooled biological replicates.

2012-02-08 | E-GEOD-27027 | biostudies-arrayexpress

Generation of thymus reconstituting T cell progenitors from genetically unmodified human pluripotent stem cells

Project description:Derivation of transplantable T cell progenitors from human pluripotent stem cells is the key step towards regenerative therapy of hereditary and acquired immunodeficiencies. Here, we describe an original methodology for differentiation of genetically non-modified human pluripotent stem cells (hPSCs) that yields T cell progenitors capable for direct and unmediated reconstitution of mouse thymus. The hPSC-derived T cell progenitors exhibited a strong thymus and spleen homing potential and developed into single positive human T cells that could enter circulation. Detailed single cell transcriptome analysis confirmed the similarity of hPSC-T cell progenitors with their in vivo counterparts. The transcription profiling attested the emergence of cells that displayed the transcription signature of early T cell progenitors.

2023-05-04 | GSE202253 | GEO

Differentiated human stem cells resemble fetal, not adult, β cells

Project description:Human pluripotent stem cells (hPSCs) have the potential to generate any human cell type, and one widely recognized goal is to make pancreatic β cells. To this end, comparisons between differentiated cell types produced in vitro and their in vivo counterparts are essential to validate hPSC-derived cells. Genome-wide transcriptional analysis of sorted insulin-expressing (INS(+)) cells derived from three independent hPSC lines, human fetal pancreata, and adult human islets points to two major conclusions: (i) Different hPSC lines produce highly similar INS(+) cells and (ii) hPSC-derived INS(+) (hPSC-INS(+)) cells more closely resemble human fetal β cells than adult β cells. This study provides a direct comparison of transcriptional programs between pure hPSC-INS(+) cells and true β cells and provides a catalog of genes whose manipulation may convert hPSC-INS(+) cells into functional β cells

2014-03-25 | GSE56130 | GEO

Differentiated human stem cells resemble fetal, not adult, Î² cells

Project description:Human pluripotent stem cells (hPSCs) have the potential to generate any human cell type, and one widely recognized goal is to make pancreatic Î² cells. To this end, comparisons between differentiated cell types produced in vitro and their in vivo counterparts are essential to validate hPSC-derived cells. Genome-wide transcriptional analysis of sorted insulin-expressing (INS(+)) cells derived from three independent hPSC lines, human fetal pancreata, and adult human islets points to two major conclusions: (i) Different hPSC lines produce highly similar INS(+) cells and (ii) hPSC-derived INS(+) (hPSC-INS(+)) cells more closely resemble human fetal Î² cells than adult Î² cells. This study provides a direct comparison of transcriptional programs between pure hPSC-INS(+) cells and true Î² cells and provides a catalog of genes whose manipulation may convert hPSC-INS(+) cells into functional Î² cells RNA is isolated and processed using MARIS from the following samples: H1 human embryonic stem cells (hESCs) in duplicate, HUES8 hESCs in duplicate, human induced pluripotent stem cells (hiPSCs) in duplicate, H1 cells differentiated to a stage in which insulin-expressing cells are present (stage 6) in duplicate, HUES8 cells differentiated to stage 6 in duplicate, hiPSCs differentiated to stage 6, insulin-expressing cells sorted from H1 cells differentiated to stage 6 in duplicate, insulin-expressing cells sorted from HUES8 cells differentiated to stage 6 in duplicate, insulin-expressing cells sorted from hiPSCs differentiated to stage 6 in duplicate, human week 16 fetal pancreata in duplicate, insulin-expressing cells sorted from human week 16 fetal pancreata in triplicate, adult human pancreatic islets in triplicate, and insulin-expressing cells sorted from adult human pancreatic islets in triplicate.

2014-03-25 | E-GEOD-56130 | biostudies-arrayexpress

Mass spectrometry-based analysis of Muse cells from different origins

Project description:Multilineage-differentiating stress enduring (Muse) cells are nontumorigenic endogenous pluripotent-like stem cells easily collected from various adult or fetal tissues. The tissue regenerative effects of Muse cells have been demonstrated in many disease models, as they reach damaged sites after intravenous injection to exert pleiotropic effects. Previous reports indicate that several human tissues are readily accessible for Muse cell isolation, including adult tissues such as bone marrow (BM) and embryonic tissues such as Wharton’s Jelly (WJ) from umbilical cord. Wa analyzed the protein repertoires of WJ-Muse and BM-Muse using mass spectrometry-based proteomics.

2023-07-17 | PXD036883 | Pride

A whole genome analysis of single fetal human stem cells from the liver and the intestine

Project description:Mutations of embryonic and fetal origin have the potential to affect a large proportion of adult cells and may alter cancer predisposition or lead to genetic disease syndromes. We have recently shown that human adult-stem cells progressively acquire approximately 40 novel tissue-specific mutations per year throughout postnatal life. Prenatal mutation rates are as yet unknown. Here we determined genome-wide mutation patterns of single stem cells in human development by sequencing of clonally expanded intestinal and liver organoid cultures of 2nd trimester human foetuses. Our results show that mutation rates in fetal stem cells are significantly higher than in adult stem cells.

| EGAS00001002886 | EGA

Quantitative analysis of redox proteome reveals protein thiol switches regulating fundamental processes in developmental hematopoiesis

Project description:Fetal and adult hematopoietic stem and progenitor cells (HSPCs) are characterized by distinct redox homeostasis that may influence their differential cellular behaviour in normal and malignant haematopoiesis. In this work, we have applied a quantitative mass spectrometry-based redox proteomic approach to comprehensively describe reversible cysteine modifications in primary mouse fetal and adult HSPCs. We defined the redox state of 4455 cysteines in fetal and adult HSPCs and demonstrated a higher susceptibility to oxidation of protein thiols in fetal HSPCs. Our data identified ontogenically active redox switches in proteins with a pronounced role in metabolism and protein homeostasis. Additional redox proteomic analysis identified redox switches acting in mitochondrial respiration as well as protein homeostasis to be triggered during onset of MLL-ENL leukemogenesis in fetal HSPCs. Our data has demonstrated that redox signalling contributes to the regulation of fundamental processes of developmental hematopoiesis and has pinpointed potential targetable redox-sensitive proteins in in utero-initiated MLL-rearranged leukaemia. An H9 human embryonic stem cells cell line was applied to validate data from the primary cells.

2022-06-09 | PXD033115 | Pride

EGR1 regulation in human bone marrow stromal cells

Project description:Human bone marrow stromal cells (BMSCs) are key elements of the hematopoietic environment and they play a central role in bone and bone marrow physiology. However, how key BMSC functions are regulated is largely unknown. We analyzed the role of the immediate early response transcription factor EGR1 as key BMSC regulator and found that EGR1 was highly expressed in prospectively-isolated primary BMSCs, downregulated upon culture, and lower in non-CFU-F-containing CD45neg BM cells. Furthermore, EGR1 expression was lower in proliferative regenerating adult and fetal primary cells compared to adult steady-state BMSCs. Accordingly, EGR1 overexpression markedly decreased BMSC proliferation but considerably improved hematopoietic stroma support function as indicated by an increased production of transplantable CD34+CD90+ hematopoietic stem cells in expansion co-cultures. The improvement of BMSC stroma support function was mediated by increased expression of hematopoietic supporting genes, such as VCAM1 and CCL28. On the other hand, EGR1 knockdown increased ROS-mediated BMSC proliferation, and clearly reduced BMSC hematopoietic stroma support potential. These findings thus show that EGR1 is a key BMSC transcription factor with a dual role in regulating proliferation and hematopoietic stroma support function that is controlling a genetic program to coordinate the specific functions of BMSC in their different biological contexts.

2019-06-27 | PXD011767 | Pride

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