Flow Cytometry and Lineage Tracing Study for Identification of Adipocyte Precursor Cell (APC) Populations

2018 ◽  
pp. 111-121 ◽  
Author(s):  
Ju Hee Lee ◽  
Azadeh Yeganeh ◽  
Hisato Konoeda ◽  
Joon Ho Moon ◽  
Hoon-Ki Sung
Keyword(s):  
Flow Cytometry ◽  
Precursor Cell ◽  
Lineage Tracing ◽  
Adipocyte Precursor

scholarly journals 4369 Reprogramming of vascular smooth muscle cells to multipotent progenitor cells contributes to progression of atherosclerosis

2020 ◽  
Vol 4 (s1) ◽  
pp. 102-102
Author(s):  
Allison Milfred Dubner ◽  
Sizhao Lu ◽  
Austin Jolly ◽  
Keith Strand ◽  
Marie Mutryn ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Progenitor Cells ◽  
Coronary Arteries ◽  
Lineage Tracing ◽  
Plaque Progression ◽  
Blood Institute ◽  
National Heart ◽  
National Heart Lung

OBJECTIVES/GOALS: Our lab previously identified a population of vascular smooth muscle (SMC)-derived progenitor cells (AdvSca1-SM) which expand robustly in response to disease and can differentiate into multiple cell types. We now aim to define the role of these AdvSca1-SM cells in atherosclerotic plaque progression. METHODS/STUDY POPULATION: Goal one uses SMC lineage tracing mice and a model of atherosclerosis to track reprogramming of SMCs to AdvSca1-SM cells in the setting of disease. Arteries are analyzed using flow cytometry and immunofluorescence to quantify changes in number of mature SMCs and AdvSca1-SM cells. Goal two uses AdvSca1-SM lineage tracing mice with high cholesterol-induced atherosclerosis and plaque neovascularization. Arteries are analyzed to quantify expansion of AdvSca1-SM cells, subsequent re-differentiation into mature SMC, endothelial cells, or macrophages, and contribution to plaque neovascularization. Mechanistic findings from both goals are being investigated in diseased human coronary arteries. RESULTS/ANTICIPATED RESULTS: Flow cytometry from SMC lineage tracing mice revealed a 7- to 13-fold expansion of AdvSca1-SM cells in carotid arteries (p<0.001) and aortas (p = 0.03) after 6 weeks of western diet; no differences in macrophage numbers were observed. Additional SMC and AdvSca1-SM cell lineage tracing mice are on atherogenic diets to assess early and advanced atherosclerosis. We predict that AdvSca1-SM cells will contribute to macrophage accumulation as well as plaque neovascularization in the setting of severe atherosclerosis. Translational relevance of mechanisms driving SMC reprogramming and AdvSca1-SM cell contribution to plaque progression are being applied to studies of diseased human coronary arteries. DISCUSSION/SIGNIFICANCE OF IMPACT: Our data suggest a role for AdvSca1-SM cells in atherosclerosis. Ongoing work will clarify the mechanisms driving plaque-associated AdvSca1-SM expansion and define the ultimate fates of these cells. In vivo modulation of this process could provide the basis for future anti-atherosclerotic therapies. CONFLICT OF INTEREST DESCRIPTION: AD - CCTSI TOTTS TL1TR002533; SL - 18POST34030397 from the American Heart Association; AJ – no conflicts; KS - 1F31HL147393 from the National Heart, Lung, and Blood Institute, NIH; MM – no conflicts; RT – no conflicts; KSM – no conflicts; RAN - R01CA236222 from the National Cancer Institute, NIH, and 2018-03 from the Lungevity Foundation; and MCMW-E - R01 HL121877 from the National Heart, Lung, and Blood Institute, NIH, and 25A8679 from the Chernowitz Foundation.

scholarly journals Search for the adipocyte precursor cell and factors that promote its differentiation.

1980 ◽  
Vol 21 (6) ◽  
pp. 657-670
Author(s):  
G J Hausman ◽  
D R Campion ◽  
R J Martin
Keyword(s):  
Precursor Cell ◽  
Adipocyte Precursor

Local progenitor cells shape the neoplastic vasculature and promote brain tumor growth.

2021 ◽  
Vol 39 (15_suppl) ◽  
pp. e14044-e14044
Author(s):  
Roland Kälin ◽  
Linzhi Cai ◽  
Dongxu Zhao ◽  
Huabin Zhang ◽  
Wenlong Zhang ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Brain Tumor ◽  
Brain Tumors ◽  
Single Cell ◽  
Progenitor Cells ◽  
Cell Population ◽  
Expression Profile ◽  
Progenitor Cell ◽  
Myeloid Cells ◽  
Lineage Tracing

e14044 Background: Aggressive brain tumors like glioblastoma depend on support by their local environment. While the role of tumor-associated myeloid cells on glioblastoma progression is well-documented, we have only partial knowledge of the pathological impact of glioblastoma -parenchymal progenitor cells. Methods: We investigated the glioblastoma microenvironment with transgenic lineage-tracing models ( nestin-creER2, R26-tdTomato and sox2-creER2,R26-tdTomato), intravital imaging, single-cell transcriptomics, immunofluorescence and flow-cytometry as well as histopathology and characterized a previously unknown tumor-associated progenitor cell. In functional experiments, we studied the knockout of the transcription factor SOX2 in these tumor-associated cells. Results: Lineage-traced cells from mouse glioblastoma were obtained by flow-cytometry and single cell transcriptomes compared to established gene expression data from brain tumor parenchymal cells. The traced tumor-associated cells had a transcriptomic profile largely resembling myeloid cells and expressed microglia-/macrophage-markers on the protein-level. However, transgenic models and bone-marrow chimera revealed that the traced cells were clearly distinct from microglia or macrophages. The traced tumor associated cells with a myeloid expression profile derived from a SOX2-dependent progenitor cell. Consequently, conditional Sox2-knockout ablated the entire myeloid-like cell population. Remarkably, this tumor-associated cell population had a large impact on disease-progression causing significant reduction of glioblastoma –vascularization to 53%, changing vascular function and leading to a decrease in tumor volume to 42% as compared to controls. The myeloid-like progenitor cells were identified in human brain tumors by immunofluorescence and in scRNA-seq data. Conclusions: We identified a previously unacknowledged population of tumor-associated progenitor cells with a myeloid-like expression profile that transiently appeared during glioblastoma growth. These progenitors have strong impact on glioblastoma progression and point towards a new and promising therapeutic target in order to support anti-angiogenic regimen in glioblastoma.

Endothelin-1 suppression of rat adipocyte precursor cell differentiation in serum-free culture.

Endocrinology ◽  
1992 ◽  
Vol 130 (4) ◽  
pp. 2031-2036
Author(s):  
O Shinohara ◽  
Y Murata ◽  
M Shimizu
Keyword(s):  
Cell Differentiation ◽  
Precursor Cell ◽  
Serum Free ◽  
Endothelin 1 ◽  
Adipocyte Precursor

scholarly journals The stream of precursors that colonizes the thymus proceeds selectively through the early T lineage precursor stage of T cell development

2008 ◽  
Vol 205 (5) ◽  
pp. 1187-1199 ◽  
Author(s):  
Claudia Benz ◽  
Vera C. Martins ◽  
Freddy Radtke ◽  
Conrad C. Bleul
Keyword(s):  
T Cell ◽  
T Cell Development ◽  
Cell Lineage ◽  
Cell Types ◽  
Cell Development ◽  
Precursor Cells ◽  
Precursor Cell ◽  
Lineage Tracing ◽  
Hematopoietic Precursor ◽  
The Common

T cell development in the thymus depends on continuous colonization by hematopoietic precursors. Several distinct T cell precursors have been identified, but whether one or several independent precursor cell types maintain thymopoiesis is unclear. We have used thymus transplantation and an inducible lineage-tracing system to identify the intrathymic precursor cells among previously described thymus-homing progenitors that give rise to the T cell lineage in the thymus. Extrathymic precursors were not investigated in these studies. Both approaches show that the stream of T cell lineage precursor cells, when entering the thymus, selectively passes through the early T lineage precursor (ETP) stage. Immigrating precursor cells do not exhibit characteristics of double-negative (DN) 1c, DN1d, or DN1e stages, or of populations containing the common lymphoid precursor 2 (CLP-2) or the thymic equivalent of circulating T cell progenitors (CTPs). It remains possible that an unknown hematopoietic precursor cell or previously described extrathymic precursors with a CLP, CLP-2, or CTP phenotype feed into T cell development by circumventing known intrathymic T cell lineage progenitor cells. However, it is clear that of the known intrathymic precursors, only the ETP population contributes significant numbers of T lineage precursors to T cell development.

scholarly journals A glucocorticoid- and diet-responsive pathway toggles adipocyte precursor cell activity in vivo

2016 ◽  
Vol 9 (451) ◽  
pp. ra103-ra103 ◽  
Author(s):  
J. C. Wong ◽  
K. C. Krueger ◽  
M. J. Costa ◽  
A. Aggarwal ◽  
H. Du ◽  
...  
Keyword(s):  
Cell Activity ◽  
Precursor Cell ◽  
Adipocyte Precursor

scholarly journals Identification and characterization of a non-conventional CD45 negative perivascular macrophage population within the mouse brain.

2021 ◽  
Author(s):  
Carole Siret ◽  
Max van Lessen ◽  
Hyun-Woo Jeong ◽  
Shuaiwei Wang ◽  
Milesa Simic ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Lineage Tracing ◽  
Cerebral Vasculature ◽  
Hematopoietic Stem ◽  
Macrophage Population ◽  
Perivascular Macrophages ◽  
Similar Location ◽  
The Brain ◽  
Identification And Characterization

Abstract Perivascular macrophages (pvM) are closely associated with cerebral vasculature and play an essential role in drainage of the brain and regulation of the immune response. Here, using reporter mouse models and immunofluorescence on sections and whole brain, flow cytometry and single cell sequencing, we identify a Lyve1+ brain perivascular population lacking classical macrophage markers such as CD45 and Cx3cr1. We named the new non-conventional CD45 negative perivascular macrophages pvM2. These cells have a similar location, morphology and phagocytic function as conventional pvM. The pvM2 are not derived from hematopoietic stem cells, as they are negative in the VavtdT lineage tracing model. They increase in number after photothrombotic induced stroke established by flow cytometry and 3D immunofluorescence analysis. Since CD45 negative cells were typically excluded from macrophage studies, the presence of pvM2 has been previously missed and their role is of importance to assess in the brain disease models.

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