Single-cell Transcriptomic Profiling of Mouse Alcam-Positive Sca-1-Negative Bone Lining Cells
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE326153 | GEO | 2026/06/01
REPOSITORIES: GEO
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Project description:Single-cell Transcriptomic Profiling of Mouse Alcam-Positive Sca-1-Negative Bone Lining Cells
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Project description:The bone marrow (BM) niche is a specialized microenvironment critical for the maintenance and regulation of hematopoietic stem cells (HSCs). Different niches in the BM support HSCs, and among them we analyzed the endosteal niche. The endosteal niche is formed by Alcam+ Sca-1- osteoblasts, Alcam- Sca-1+ mesenchymal stem cells (MSCs), and Alcam- Sca-1- cells. Using single-cell gene expression analysis, we identified a unique subpopulation within the Alcam+ Sca-1- bone-lining cells (BLCs) that lacked osteoblast markers. This subpopulation was found to be a novel MSC with specific expression of the 8-integrin (Itga8). We compared gene expression between Itga8+ Alcam+ Sca-1- MSCs (Itga8+ MSCs), Itga8- Alcam+ Sca-1- BLCs, Alcam- Sca-1+ MSCs, and CD51+ CD140a+ MSCs to investigate the characteristics of Itga8+ MSCs. Itga8+ MSCs showed a distinct gene expression profile compared with the others, demonstrating that Itga8+ MSCs are different from previously reported BLCs and MSCs. Furthermore, Itga8+ MSCs were found to secrete extracellular matrix (ECM) that regulates HSCs, and therefore, Itga8+ MSCs are important MSCs that provide the ECM required to maintain HSCs within the endosteal niche.
2026-05-27 | GSE280158 | GEO
Project description:A subset of osteoblasts functions as a key component of the hematopoietic stem cell niche. But it is still unclear that the precise cellular and molecular contribution of endosteal cell populations on the HSC supportive microenvironment. We purified the endosteal cells and subdivided them into mesenchymal progenitor cells (MPCs: ALCAM(-)Sca-1(+)) and two osteoblast populations (ALCAM(-)Sca-1(-) and ALCAM(+)Sca-1(-)). In this study, to clarify the characteristics of these fractionated cells in detail, gene expression analysis were performed using the GeneChip Mouse Genome Array 430 2.0 (Affymetrix). The bone fragments of femur and tibiae were treated with collagenase and endosteal cells were obtained. For enrichment of non-hematopoietic and non-endothelial population, CD45(-)CD31(-)Ter119(-) population was isolated and then the expressions of Sca-1 and activated leukocyte cell adhesion molecule (ALCAM) was analyzed by FACS. FACS analysis showed that CD45(-)CD31(-)Ter119(-) cells were subdivided into ALCAM(-)Sca-1(+), ALCAM(-)Sca-1(-) and ALCAM(+)Sca-1(-). These three populations were sorted and RNA samples were used for amplification, labeling, and hybridization to independent arrays.
2010-08-10 | E-GEOD-17569 | biostudies-arrayexpress
Project description:Signal differences between TMRM-hi and ALCAM-low vs. TMRM-hi and ALCAM-hi in representative cardiac and hepatic gene expression in mRNA microarray analysis. INTRODUCTION: Non-genetic purification methods for pluripotent stem cell-derived hepatocyte-like cells are useful for liver regenerative therapy and pharmaceutical applications. METHODS: Fluorescent activated cell sorting (FACS) was used to separate cells by combining two parameters: cellular mitochondrial content evaluated by the mitochondrial membrane potential-dependent fluorescent probe (TMRM) and immunocytochemical detection of activated leukocyte cell adhesion molecule (ALCAM). This method was applied to murine fetal, human embryonic stem cell (ESC)-derived, and human induced pluripotent stem cell (iPSC)-derived cell-mixtures. Separately sorted cell fractions were evaluated by quantitative PCR, immunohistochemistry, and cytochemistry for HNF4a, AFP, and albumin mRNA and/or protein expression. RESULTS: Hepatocyte-like cells were highly segregated into the high TMRM signal and ALCAM-positive population. The purity of hepatocyte-like cells derived from human iPSCs was 97 ± 0.38% (n = 5). CONCLUSIONS: This hepatocyte-like cell purification method may be applicable to quality control of cells for liver regenerative cell therapy and pharmaceutical development.
2020-03-01 | GSE126812 | GEO
Project description:Endometrial cancer is the most common gynaecological cancer in western countries, being the endometrioid tumours the most common subtype. Most patients are diagnosed at an early stage and present an excellent prognosis. However, a consistent number of those continue to suffer recurrence, without means to be identified by risk classification systems. Thus, finding a reliable marker to predict recurrence becomes an important unmet clinical issue. ALCAM is a cell–cell adhesion molecule, member of the Immunoglobulin superfamily, which has been associated to the carcinogenesis of many cancers. In here, we first determined the value of ALCAM as marker of recurrence in endometrioid endometrial cancer by conducting a retrospective multicenter study in 174 primary tumours. In early stage patients (N=134), recurrence-free survival was poorer in patients with ALCAM-positive compared to ALCAM-negative tumours (HR 4.237; 95%CI 1.01-17.76). This difference was more significant in patients with early stage moderately-poorly differentiated tumours (HR 9,259; 95%CI 2.12-53.47). In multivariate analysis, ALCAM-positivity was an independent prognostic factor in early stage disease (HR 6.027, 95% CI 1.41-25.74). Then, we demonstrated in vitro a role for ALCAM in cell migration and invasion by using a loss-of-function model in two endometrial cancer cell lines. Moreover, ALCAM depletion resulted in a reduced primary tumour size and reduced metastatic local spread in an orthotopic murine model. Gene expression analysis of ALCAM-depleted cell lines supported that motility, invasiveness, cellular assembly and organization were the most deregulated functions associated to ALCAM. Finally, we evidenced some of the downstream effector genes that are involved in ALCAM mediated cell migration; specifically FLNB, TXNRD1 and LAMC2 were validated. In conclusion, our results highlight the potential of ALCAM as a recurrent biomarker in early stage endometrioid endometrial cancer and point to ALCAM as an important molecule in endometrial cancer dissemination by regulating cell migration, invasion and metastasis.
2016-09-08 | GSE86543 | GEO
Project description:Stem cells antigen-1 (Sca-1) is an 18-kDa mouse glycosyl phosphatidylinositol-anchored cell surface protein that has widely been used for isolation of murine hematopoietic stem cells (HSCs). However, recently several studies have reported the expression of this protein in non-HSCs such as the liver, skin and muscles among others. In the current study, we used fluorescence-activated cell sorting (FACS) technique to sort Sca-1+ stem cells from mouse hindlimb muscles. The Sca-1+ cells were divided into two sets first set of Sca-1+ cells were processed for protein extraction immediately after sorting (ex vivo set) while the second set of samples, referred to as in vitro set, were expanded in cell culture after sorting and harvested after two passage and then processed for protein extraction. A high-resolution proteome map of the ex vivo and in vitro sets was generated Orbitrap Fusion Tribrid Mass Spectrometer. In total, the analysis led to the identification of 5,581 protein groups.
2021-06-21 | PXD022247 | Pride
Project description:A subset of osteoblasts functions as a key component of the hematopoietic stem cell niche. But it is still unclear that the precise cellular and molecular contribution of endosteal cell populations on the HSC supportive microenvironment. We purified the endosteal cells and subdivided them into mesenchymal progenitor cells (MPCs: ALCAM(-)Sca-1(+)) and two osteoblast populations (ALCAM(-)Sca-1(-) and ALCAM(+)Sca-1(-)). In this study, to clarify the characteristics of these fractionated cells in detail, gene expression analysis were performed using the GeneChip Mouse Genome Array 430 2.0 (Affymetrix).
2010-08-10 | GSE17569 | GEO
Project description:Lining macrophages initiate articular inflammation
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Project description:This dataset contains the transcriptional analysis of lining macrophages and sub-lining macrophages from vehicle or antigen-induced arthritis(AIA) mouse model.
2023-04-10 | GSE229189 | GEO
Project description:We discovered that mice that lack Lsd1 in hematopoietic cells were exhibited increased frequencies of CD150+ CD48- lin- c-Kit+ Sca-1+ LT-HSCs, but completely lacked the lin- c-Kit+ Sca-1- myeloid progenitor compartment. To determine the genes altered by Lsd1-loss, CD150+ CD48- lin- c-Kit+ Sca-1+ LT-HSCs from Lsd1fl/fl and Lsd1fl/fl Mx1Cre mice were FACS-purified to be analyzed by gene expression profiling.
2013-06-25 | GSE40284 | GEO