scholarly journals Generation of Dispersed Presomitic Mesoderm Cell Cultures for Imaging of the Zebrafish Segmentation Clock in Single Cells

10.3791/50307 ◽  
2014 ◽  
Author(s):  
Alexis B. Webb ◽  
Daniele Soroldoni ◽  
Annelie Oswald ◽  
Johannes Schindelin ◽  
Andrew C. Oates
Keyword(s):  
Cell Cultures ◽  
Single Cells ◽  
Presomitic Mesoderm ◽  
Mesoderm Cell ◽  
Segmentation Clock

Effects of carbamylcholine on chick embryo aggregate retina cell cultures

1988 ◽  
Vol 46 ◽  
pp. 350-351
Author(s):  
Glenn M. Cohen ◽  
Radharaman Ray
Keyword(s):  
Cell Cultures ◽  
Single Cells ◽  
Retinal Cell ◽  
Cell Aggregates ◽  
High Concentration ◽  
In Ovo ◽  
Sterile Culture ◽  
Retinal Tissue ◽  
Synaptic Junctions ◽  
And Control

Retinal,cell aggregates develop in culture in a pattern similar to the in ovo retina, forming neurites first and then synapses. In the present study, we continuously exposed chick retinal cell aggregates to a high concentration (1 mM) of carbamylcholine (carbachol), an acetylcholine (ACh) analog that resists hydrolysis by acetylcholinesterase (AChE). This situation is similar to organophosphorus anticholinesterase poisoning in which the ACh level is elevated at synaptic junctions due to inhibition of AChE, Our objective was to determine whether continuous carbachol exposure either damaged cholino- ceptive neurites, cell bodies, and synaptic elements of the aggregates or influenced (hastened or retarded) their development.The retinal tissue was isolated aseptically from 11 day embryonic White Leghorn chicks and then enzymatically (trypsin) and mechanically (trituration) dissociated into single cells. After washing the cells by repeated suspension and low (about 200 x G) centrifugation twice, aggregate cell cultures (about l0 cells/culture) were initiated in 1.5 ml medium (BME, GIBCO) in 35 mm sterile culture dishes and maintained as experimental (containing 10-3 M carbachol) and control specimens.

scholarly journals Colony formation of clone-sorted human hematopoietic progenitors

Blood ◽  
1990 ◽  
Vol 75 (10) ◽  
pp. 1941-1946 ◽  
Author(s):  
H Ema ◽  
T Suda ◽  
Y Miura ◽  
H Nakauchi
Keyword(s):  
Single Cell ◽  
Cell Cultures ◽  
Mononuclear Cells ◽  
Direct Action ◽  
Single Cells ◽  
Accurate Estimation ◽  
Hematopoietic Progenitors ◽  
Gm Csf ◽  
Cd34 Cells ◽  
Sorting System

Abstract To characterize human hematopoietic progenitors, we performed methylcellulose cultures of single cells isolated from a population of CD34+ cells by fluorescence-activated cell-sorting (FACS) clone-sorting system. CD34+ cells were detected in bone marrow (BM) and peripheral blood (PB) cells at incidences of 1.0% and 0.01% of total mononuclear cells, respectively. Single cell cultures revealed that approximately 37% of BM CD34+ cells formed colonies in the presence of phytohemagglutinin-leukocyte conditioned medium and erythropoietin. Erythroid bursts-, granulocyte-macrophage (GM) colony-, and pure macrophage (Mac) colony-forming cells were 10% each in CD34+ cells. Approximately 15% of PB CD34+ cells formed colonies in which erythroid bursts were predominant. CD34+ cells were heterogeneous and fractionated by several antibodies in FACS multicolor analysis. In these fractionated cells, CD34+, CD33+ cells formed GM and Mac colonies 7 to 10 times as often as CD34+, CD33- cells. Most of the erythroid bursts and colonies were observed in the fraction of CD34+, CD13- cells or CD34+, CD33- cells. The expression of HLA-DR on CD34+ cells was not related to the incidence, size, or type of colonies. There was no difference in the phenotypical heterogeneity of CD34+ cells between BM and PB. About 10% of CD34+ cells were able to form G colonies in response to granulocyte colony-stimulating factor (G-CSF) and to form Mac colonies in GM-CSF or interleukin-3 (IL-3). Progenitors capable of generating colonies by stimulation of G-CSF were more enriched in CD34+, CD33+ fraction than in CD34+, CD33- fraction. Thus, single cell cultures using the FACS clone-sorting system provide an accurate estimation of hematopoietic progenitors and an assay system for direct action of colony-stimulating factors.

Modeling of segmentation clock mechanism in presomitic mesoderm

2009 ◽  
Author(s):  
A. Kazama ◽  
A. Karashima ◽  
N. Katayama ◽  
M. Nakao
Keyword(s):  
Presomitic Mesoderm ◽  
Segmentation Clock ◽  
Clock Mechanism

scholarly journals STUDIES ON THE MODE OF SPREAD OF B. ENTERITIDIS MOUSE TYPHOID INFECTION

1927 ◽  
Vol 46 (6) ◽  
pp. 871-886 ◽  
Author(s):  
Leslie T. Webster ◽  
Caspar Burn
Keyword(s):  
Growth Rate ◽  
Single Cell ◽  
Cell Cultures ◽  
Bacterial Cell ◽  
Single Cells ◽  
Oxygen Absorption ◽  
Acid Solutions ◽  
Staining Property ◽  
Low Degree ◽  
Cell Changes

1. During early stages of multiplication, single cells from smooth-, mucoid-, and rough-susceptible and variant colonies show no differences in morphology or growth rate. 2. Cells from 18 to 24 hour single cell cultures of these various colony types possess similar oxygen absorption and cataphoretic migratory rates. In staining property, the cells from mucoid colonies appear larger, and those from rough colonies smaller, than the typical cells from smooth-susceptible colonies. 3. Cells from bacteriophage-resistant colonies differ from those of bacteriophage-susceptible colonies in their ability to multiply luxuriantly in the presence of bacteriophage, and in their tendency to flocculate in acid solutions at pH 3.8 to 4.1, as well as in their low degree of virulence. 4. Cells from smooth bacteriophage-susceptible colonies in contact with bacteriophage under conditions where multiplication is restrained may be altered so as to resemble the cells from the bacteriophage-resistant colonies. 5. These facts furnish evidence that bacteriophage adheres to the surface of the bacterial cell and that the various cell changes and colony alterations are of an environmental rather than genetic nature.

scholarly journals Murine Peyer's patch T cell clones. Characterization of antigen-specific helper T cells for immunoglobulin A responses.

1982 ◽  
Vol 156 (4) ◽  
pp. 1115-1130 ◽  
Author(s):  
H Kiyono ◽  
J R McGhee ◽  
L M Mosteller ◽  
J H Eldridge ◽  
W J Koopman ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
B Cell ◽  
Cell Cultures ◽  
Single Cells ◽  
Immunoglobulin A ◽  
Anatomical Site ◽  
Continuous Growth ◽  
A Cells ◽  
Helper Activity

We successfully cloned antigen-specific T cells from murine gut-associated lymphoreticular tissue, i.e., Peyer's patches, which are dependent upon T cell growth factor and independent of antigen for continuous growth. These clones exhibit helper activity for IgA responses to sheep erythrocytes (SRBC) and have been designated T helper (Th) A. Two broad categories of Th A clones have been maintained in continuous culture. The first group supports IgM and largely IgA anti-SRBC plaque-forming cell (PFC) responses in both normal and SRBC-primed splenic B cell cultures, whereas the second group supports low IgM, IgG1, and IgG2 and high IgA PFC responses. Subclones derived from single cells maintain the parent helper properties when propagated in culture for long periods (greater than 7 mo). Cloned Th A cells are antigen specific and do not support polyclonal or immune responses to other thymus dependent antigens in normal B cell cultures. Th A cells require full histocompatibility for helper functions because addition of cloned Th A cells to B cell cultures from other H-2 types does not result in IgA responses. Cloned Th A cells are Thy-1.2+ and Lyt-1+ and Lyt-2-, Ig-, and I-A-. Th A cells bear Fc receptors for IgA and do not possess receptors for IgM or IgG isotypes. Thus, T cells that primarily promote IgA isotype responses have been isolated in high frequency from murine PP, an anatomical site of major importance for induction and regulation of the IgA response.

scholarly journals Deep learning guided image-based droplet sorting for on-demand selection and analysis of single cells and 3D cell cultures

Lab on a Chip ◽  
2020 ◽  
Vol 20 (5) ◽  
pp. 889-900 ◽  
Author(s):  
Vasileios Anagnostidis ◽  
Benjamin Sherlock ◽  
Jeremy Metz ◽  
Philip Mair ◽  
Florian Hollfelder ◽  
...  
Keyword(s):  
Neural Networks ◽  
Deep Learning ◽  
Cell Cultures ◽  
Mammalian Cells ◽  
Single Cells ◽  
Bright Field ◽  
On Demand ◽  
3D Cell Cultures ◽  
Droplet Sorting ◽  
Demand Selection

To uncover the heterogeneity of cellular populations and multicellular constructs we show on-demand isolation of single mammalian cells and 3D cell cultures by coupling bright-field microdroplet imaging with real-time classification and sorting using convolutional neural networks.

Cytogenetic abnormalities in cotton (Gossypium hirsutum L.) cell cultures

Genome ◽  
1989 ◽  
Vol 32 (6) ◽  
pp. 1128-1134 ◽  
Author(s):  
Ruzhong Li ◽  
David M. Stelly ◽  
Norma L. Trolinder
Keyword(s):  
Gossypium Hirsutum ◽  
Cell Cultures ◽  
Single Cells ◽  
Metaphase Chromosomes ◽  
Gossypium Hirsutum L ◽  
Low Frequencies ◽  
Cytogenetic Abnormalities ◽  
Paired Samples ◽  
Ring Chromosomes

High frequencies of somaclonal phenotypic and cytogenetic variation have been observed previously among regenerants from cotton (Gossypium hirsutum L., 2n = 4x = 52). In this study we endeavored to determine if cytogenetic abnormalities would be detectable in cotton cell cultures and if so, whether or not the observed abnormalities would parallel those expected on the basis of previous cytogenetic analyses of cotton somaclones. Paired samples from suspension cultures established from 21-month-old 'Coker 312' and 8-month-old 'Coker 315' calli were pretreated or not pretreated with colchicine to detect cytogenetic abnormalities at metaphase or anaphase–telophase, respectively. Cell cultures established from both calli were found to vary in chromosome number. Hypoaneuploidy was common, but hyperaneuploidy and polyploidy were rare. Modal chromosome numbers for the 'Coker 312' and 'Coker 315' cultures were 46 and 50, respectively. Bridges at anaphase and telophase were frequent in the 'Coker 312' cultures but rare in the 'Coker 315'cultures. Cytogenetic differences between the cultures could be due to effects of culture age, genotype, their interaction, or other factors. Very small chromosomes, presumably centric fragments, as well as ring chromosomes and putative bridges between metaphase chromosomes occurred at low frequencies. The prevalence of hypoaneuploidy and rarity of hyperaneuploidy and polyploidy in cultures paralleled previous results on cotton somaclones, indicating that cytogenetic abnormalities arising in vitro probably contribute significantly to cotton somaclonal variation. The occurrence of hypoaneuploidy and bridges, including multiple bridges within single cells, is concordant with the hypothesis that breakage–fusion–bridge cycles may accumulate during in vitro culture of cotton.Key words: cotton, Gossypium, tissue culture, cytogenetics.

scholarly journals Segmentation-clock synchronization in circular-lattice networks of embryonic presomitic-mesoderm cells

2021 ◽  
Vol 6 (6) ◽  
pp. 5817-5836
Author(s):  
Jesús Pantoja-Hernández ◽  
◽  
Moisés Santillán
Keyword(s):  
Clock Synchronization ◽  
Presomitic Mesoderm ◽  
Segmentation Clock

scholarly journals Axial skeletal defects caused by mutation in the spondylocostal dysplasia/pudgy geneDll3are associated with disruption of the segmentation clock within the presomitic mesoderm

Development ◽  
2002 ◽  
Vol 129 (7) ◽  
pp. 1795-1806 ◽  
Author(s):  
Sally L. Dunwoodie ◽  
Melanie Clements ◽  
Duncan B. Sparrow ◽  
Xin Sa ◽  
Ronald A. Conlon ◽  
...  
Keyword(s):  
Gene Targeting ◽  
Skeletal Dysplasia ◽  
Null Allele ◽  
Developmental Origins ◽  
Loss Of Function ◽  
Presomitic Mesoderm ◽  
Phenotypic Analysis ◽  
Segmentation Clock ◽  
Skeletal Defects ◽  
Somite Formation

A loss-of-function mutation in the mouse delta-like3 (Dll3) gene has been generated following gene targeting, and results in severe axial skeletal defects. These defects, which consist of highly disorganised vertebrae and costal defects, are similar to those associated with the Dll3-dependent pudgy mutant in mouse and with spondylocostal dysplasia (MIM 277300) in humans. This study demonstrates that Dll3neo and Dll3pu are functionally equivalent alleles with respect to the skeletal dysplasia, and we suggest that the three human DLL3 mutations associated with spondylocostal dysplasia are also functionally equivalent to the Dll3neo null allele. Our phenotypic analysis of Dll3neo/Dll3neo mutants shows that the developmental origins of the skeletal defects lie in delayed and irregular somite formation, which results in the perturbation of anteroposterior somite polarity. As the expression of Lfng, Hes1, Hes5 and Hey1 is disrupted in the presomitic mesoderm, we suggest that the somitic aberrations are founded in the disruption of the segmentation clock that intrinsically oscillates within presomitic mesoderm.

scholarly journals PAPC couples the Segmentation Clock to somite morphogenesis by regulating N-cadherin dependent adhesion

2016 ◽  
Author(s):  
Jerome Chal ◽  
Charlene Guillot ◽  
Olivier Pourquie
Keyword(s):  
Mouse Embryos ◽  
Loss Of Function ◽  
Presomitic Mesoderm ◽  
Segmentation Clock ◽  
Anterior Compartment ◽  
Chicken Embryos ◽  
Differential Adhesion ◽  
Complementary Pattern ◽  
The Future

Vertebrate segmentation is characterized by the periodic formation of epithelial somites from the mesenchymal presomitic mesoderm (PSM). How the rhythmic signaling pulse delivered by the Segmentation Clock is translated into the periodic morphogenesis of somites remains poorly understood. Here, we focused on the role of Paraxial protocadherin (PAPC/Pcdh8) in this process. We show that in chicken and mouse embryos, PAPC expression is tightly regulated by the Clock and Wavefront system in the posterior PSM. We observed that PAPC exhibits a striking complementary pattern to N-Cadherin (CDH2), marking the interface of the future somite boundary in the anterior PSM. Gain and loss of function of PAPC in chicken embryos disrupt somite segmentation by altering the CDH2-dependent epithelialization of PSM cells. Our data suggest that clathrin-mediated endocytosis is increased in PAPC expressing cells, subsequently affecting CDH2 internalization in the anterior compartment of the future somite. This in turn generates a differential adhesion interface, allowing formation of the acellular fissure that defines the somite boundary. Thus periodic expression of PAPC downstream of the Segmentation Clock triggers rhythmic endocytosis of CDH2, allowing for segmental de-adhesion and individualization of somites.

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