Isolation and characterization of porcine visceral endoderm cell lines derived from in vivo 11-day blastocysts

Study Design. Isolation and characterization of human epidural fat (HEF) stem/progenitor cells. Objective. To identify a progenitor population within HEF and to determine if they meet the minimal criteria of a mesenchymal stem cell (MSC). Summary of Background Data. The biological function, if any, has yet to be determined for HEF. The presence of MSCs within HEF may indicate a regenerative potential within the HEF. Methods. HEF was isolated from 10 patients during elective spinal surgery. HEF cells were differentiated along osteo-, adipo-, and chondrogenic lineages, with differentiation analyzed via qPCR and histology. The cell surface receptor profile of HEF cells was examined by flow cytometry. HEF cells were also assayed through the collagen contraction assay. Prx1CreERT2GFP:R26RTdTomato MSC lineage-tracking mice were employed to identify EF MSCs in vivo. Results. HEF cell lines were obtained from all 10 patients in the study. Cells from 2/10 patients demonstrated full MSC potential, while cells from 6/10 patients demonstrated progenitor potential; 2/10 patients presented with cells that retained only adipogenic potential. HEF cells demonstrated MSC surface marker expression. All patient cell lines contracted collagen gels. A Prx1-positive population in mouse epidural fat that appeared to contribute to the dura of the spinal cord was observed in vivo. Conclusions. MSC and progenitor populations are present within HEF. MSCs were not identified in all patients examined in the current study. Furthermore, all patient lines demonstrated collagen contraction capacity, suggesting either a contaminating activated fibroblast population or HEF MSCs/progenitors also demonstrating a fibroblast-like phenotype. In vivo analysis suggests that these cell populations may contribute to the dura. Overall, these results suggest that cells within epidural fat may play a biological role within the local environment above providing a mechanical buffer.

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Characterization of stem cell–like cancer cells in immune-competent mice

Blood ◽

10.1182/blood-2006-05-024687 ◽

2006 ◽

Vol 108 (12) ◽

pp. 3906-3912 ◽

Cited By ~ 63

Author(s):

Jorg A. Kruger ◽

Charles D. Kaplan ◽

Yunping Luo ◽

He Zhou ◽

Dorothy Markowitz ◽

...

Keyword(s):

Stem Cell ◽

Cancer Cells ◽

Cell Lines ◽

Carcinoma Cell ◽

Side Population ◽

Carcinoma Cell Lines ◽

Tumorigenic Potential ◽

4T1 Cells

AbstractRecently, the cancer stem cell hypothesis has gained significant recognition as the descriptor of tumorigenesis. Although previous studies relied on transplanting human or rat tumor cells into immunecompromised mice, our study used the Hoechst 33342 dye–based side population (SP) technique to isolate and transplant stem cell–like cancer cells (SCLCCs) from the 4T1 and NXS2 murine carcinoma cell lines into the immune-competent microenvironment of syngeneic mice. 4T1 cells displayed an SP of 2% with a Sca-1highc-Kit–CD45– phenotype, whereas NXS2 cells contained an SP of 0.2% with a Sca-1highCD24highc-Kit–CD45–GD high2 phenotype. Reverse transcription–polymerase chain reaction (RT-PCR) further revealed up-regulation in SP cells of ABCG2, Sca-1, Wnt-1, and TGF-β2. Additionally, 4T1 and NXS2 SP cells exhibited increased resistance to chemotherapy, and 4T1 SP cells also showed an increased ability to efflux doxorubicin, which correlated with a selective increase in the percentage of SP cells found in the tumors of doxorubicin-treated mice. Most importantly, SP cells showed a markedly higher repopulation and tumorigenic potential in vivo, which correlated with an increased number of cells in the SP compartment of SP-derived tumors. Taken together, these results show that we successfully characterized SCLCCs from 2 murine carcinoma cell lines in the immune-competent microenvironment of syngeneic mice.

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Isolation and characterization of conditionally immortalized astrocyte cell lines derived from adult human spinal cord

Glia ◽

10.1002/glia.440100308 ◽

1994 ◽

Vol 10 (3) ◽

pp. 211-226 ◽

Cited By ~ 39

Author(s):

Scott R. Whittemore ◽

Joseph T. Neary ◽

Naomi Kleitman ◽

Henry R. Sanon ◽

Adelaida Benigno ◽

...

Keyword(s):

Spinal Cord ◽

Cell Lines ◽

Human Spinal Cord ◽

Adult Human ◽

Isolation And Characterization

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Isolation and characterization of a gene (CBF2) specifying a protein component of the budding yeast kinetochore.

The Journal of Cell Biology ◽

10.1083/jcb.121.3.513 ◽

1993 ◽

Vol 121 (3) ◽

pp. 513-519 ◽

Cited By ~ 63

Author(s):

W Jiang ◽

J Lechner ◽

J Carbon

Keyword(s):

Molecular Motors ◽

Cell Biology ◽

Budding Yeast ◽

Isolation And Characterization ◽

Sequence Homologies ◽

Binding Factor ◽

Microtubule Motor

We have cloned and determined the nucleotide sequence of the gene (CBF2) specifying the large (110 kD) subunit of the 240-kD multisubunit yeast centromere binding factor CBF3, which binds selectively in vitro to yeast centromere DNA and contains a minus end-directed microtubule motor activity. The deduced amino acid sequence of CBF2p shows no sequence homologies with known molecular motors, although a consensus nucleotide binding site is present. The CBF2 gene is essential for viability of yeast and is identical to NDC10, in which a conditional mutation leads to a defect in chromosome segregation (Goh, P.-Y., and J. V. Kilmartin, in this issue of The Journal of Cell Biology). The combined in vitro and in vivo evidence indicate that CBF2p is a key component of the budding yeast kinetochore.

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Isolation and characterization of adrenocortical progenitors involved in the adaptation to stress

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1814072115 ◽

2018 ◽

Vol 115 (51) ◽

pp. 12997-13002 ◽

Cited By ~ 15

Author(s):

Charlotte Steenblock ◽

Maria F. Rubin de Celis ◽

Luis F. Delgadillo Silva ◽

Verena Pawolski ◽

Ana Brennand ◽

...

Keyword(s):

Lineage Tracing ◽

Differentiated Cells ◽

Isolation And Characterization ◽

Proliferation And Differentiation ◽

Response To Stress ◽

Steroidogenic Cells ◽

High Degree

The adrenal gland is a master regulator of the human body during response to stress. This organ shows constant replacement of senescent cells by newly differentiated cells. A high degree of plasticity is critical to sustain homeostasis under different physiological demands. This is achieved in part through proliferation and differentiation of adult adrenal progenitors. Here, we report the isolation and characterization of a Nestin+ population of adrenocortical progenitors located under the adrenal capsule and scattered throughout the cortex. These cells are interconnected with progenitors in the medulla. In vivo lineage tracing revealed that, under basal conditions, this population is noncommitted and slowly migrates centripetally. Under stress, this migration is greatly enhanced, and the cells differentiate into steroidogenic cells. Nestin+ cells cultured in vitro also show multipotency, as they differentiate into mineralocorticoid and glucocorticoid-producing cells, which can be further influenced by the exposure to Angiotensin II, adrenocorticotropic hormone, and the agonist of luteinizing hormone-releasing hormone, triptorelin. Taken together, Nestin+ cells in the adult adrenal cortex exhibit the features of adrenocortical progenitor cells. Our study provides evidence for a role of Nestin+ cells in organ homeostasis and emphasizes their role under stress. This cell population might be a potential source of cell replacement for the treatment of adrenal insufficiency.

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Pharmaceutical properties of 'sucupira' (Pterodon spp.)

Brazilian Journal of Pharmaceutical Sciences ◽

10.1590/s1984-82502010000400002 ◽

2010 ◽

Vol 46 (4) ◽

pp. 607-616 ◽

Cited By ~ 15

Author(s):

Daiane Hansen ◽

Mitsue Haraguchi ◽

Antonio Alonso

Keyword(s):

Biological Effects ◽

Chemical Compounds ◽

Experimental Models ◽

Popular Medicine ◽

Isolation And Characterization ◽

Pharmaceutical Properties ◽

The Relationship

The plant of the genus Pterodon (Fabaceae, Leguminosae), commonly known as 'sucupira' or 'faveira', are disseminated throughout the central region of Brazil and has frequently been used in popular medicine for its anti-rheumatic, analgesic, and anti-inflammatory properties. In recent years, interest in these plants has increased considerably. The biological effects of different phytoextracts and pure metabolites have been investigated in several experimental models in vivo and in vitro. The literature describes flavonoids, triterpene and steroids, while one paper presented studies with proteins isolated from the genus. This review provides an overview of phytochemical and pharmacological research in Pterodon, showing the main chemical compounds studied to date, and focusing on the relationship between these molecules and their biological activity. Furthermore, this study paves the way for more in-depth investigation, isolation and characterization of the molecules of this plant genus.

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Novel approaches toward inhibition of the Maillard reaction in vivo: Search, isolation and characterization of prokaryotic enzymes which degrade glycated substrates

Oxidative Stress and Aging ◽

10.1007/978-3-0348-7337-6_15 ◽

1995 ◽

pp. 141-149 ◽

Cited By ~ 10

Author(s):

V. M. Monnier ◽

C. Gerhardinger ◽

M. S. Marion ◽

S. Taneda

Keyword(s):

Maillard Reaction ◽

Isolation And Characterization ◽

Novel Approaches

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Isolation and characterization of the dopa decarboxylase gene of Drosophila melanogaster

Molecular and Cellular Biology ◽

10.1128/mcb.1.6.475-485.1981 ◽

1981 ◽

Vol 1 (6) ◽

pp. 475-485

Author(s):

J Hirsh ◽

N Davidson

Keyword(s):

Drosophila Melanogaster ◽

Ribonucleic Acid ◽

Deoxyribonucleic Acid ◽

Isolation Procedure ◽

Chromosome Band ◽

Dopa Decarboxylase ◽

Developmental Profile ◽

Isolation And Characterization

We have isolated chromosomal deoxyribonucleic acid clones containing the Drosophila dopa decarboxylase gene. We describe an isolation procedure which can be applied to other nonabundantly expressed Drosophila genes. The dopa decarboxylase gene lies within or very near polytene chromosome band 37C1-2. The gene is interrupted by at least one intron, and the primary mode of regulation is pretranslational. At least two additional sequences hybridized by in vivo ribonucleic acid-derived probes are found within a 35-kilobase region surrounding the gene. The developmental profile of ribonucleic acid transcribed from one of these regions differs from that of the dopa decarboxylase transcript.

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Ex Vivo Isolation and Characterization of Cd4+Cd25+ T Cells with Regulatory Properties from Human Blood

Journal of Experimental Medicine ◽

10.1084/jem.193.11.1303 ◽

2001 ◽

Vol 193 (11) ◽

pp. 1303-1310 ◽

Cited By ~ 797

Author(s):

Detlef Dieckmann ◽

Heidi Plottner ◽

Susanne Berchtold ◽

Thomas Berger ◽

Gerold Schuler

Keyword(s):

T Cells ◽

Cytotoxic T Lymphocyte ◽

Ex Vivo ◽

Cell Receptor ◽

Regulatory Cells ◽

Suppressor T Cells ◽

Isolation And Characterization ◽

Regulatory Properties

It has been known for years that rodents harbor a unique population of CD4+CD25+ “professional” regulatory/suppressor T cells that is crucial for the prevention of spontaneous autoimmune diseases. Here we demonstrate that CD4+CD25+CD45RO+ T cells (mean 6% of CD4+ T cells) are present in the blood of adult healthy volunteers. In contrast to previous reports, these CD4+CD25+ T cells do not constitute conventional memory cells but rather regulatory cells exhibiting properties identical to their rodent counterparts. Cytotoxic T lymphocyte–associated antigen (CTLA)-4 (CD152), for example, which is essential for the in vivo suppressive activity of CD4+CD25+ T cells, was constitutively expressed, and remained strongly upregulated after stimulation. The cells were nonproliferative to stimulation via their T cell receptor for antigen, but the anergic state was partially reversed by interleukin (IL)-2 and IL-15. Upon stimulation with allogeneic (but not syngeneic) mature dendritic cells or platebound anti-CD3 plus anti-CD28 the CD4+CD25+ T cells released IL-10, and in coculture experiments suppressed the activation and proliferation of CD4+ and CD8+ T cells. Suppression proved IL-10 independent, yet contact dependent as in the mouse. The identification of regulatory CD4+CD25+ T cells has important implications for the study of tolerance in man, notably in the context of autoimmunity, transplantation, and cancer.

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