Dissociation of insect Malpighian tubules into single, viable cells

1984 ◽  
Vol 72 (1) ◽  
pp. 101-109
Author(s):  
W.M. Satmary ◽  
T.J. Bradley
Keyword(s):  
Single Cells ◽  
Stellate Cells ◽  
Malpighian Tubules ◽  
Cell Type ◽  
Isolated Cells ◽  
Trypan Blue Exclusion ◽  
Distinct Cell Type ◽  
Viable Cells ◽  
Distinct Cell ◽  
Aedes Taeniorhynchus

The Malpighian tubules of insects are generally composed of more than one cell type. In the hemipteran Rhodnius prolixus, the tubules are divided into two regions, termed the upper and lower tubules, each of which is composed of a distinct cell type. In the dipteran Aedes taeniorhynchus, primary and stellate cells are interspersed throughout the length of the tubules. We report here techniques for the dissociation of the Malpighian tubules of both of these species into single cells. Tubules are removed from the insect and placed for 1 h in insect Ringer containing elastase (Sigma, type III) at 4 mg/ml. This treatment fully removes the basal lamella. Mild agitation by hand produces a suspension of single cells, which remain viable as determined by Trypan Blue exclusion. Isolated cells have been maintained in cell culture for one week. Using light and scanning electron microscopy, upper and lower tubule cells of Rhodnius and primary and stellate cells of Aedes can be distinguished on the basis of size, shape, microvillar length, and the presence or absence of intracellular crystals.

Isolation of smooth muscle cells from swine carotid artery by digestion with papain

1986 ◽  
Vol 251 (3) ◽  
pp. C474-C481 ◽  
Author(s):  
S. P. Driska ◽  
R. Porter
Keyword(s):  
Smooth Muscle ◽  
Carotid Artery ◽  
Smooth Muscle Cells ◽  
Single Cells ◽  
Calcium Ionophore ◽  
Muscle Cells ◽  
Ionophore A23187 ◽  
Free Solution ◽  
Isolated Cells ◽  
Viable Cells

A new method is described for the preparation of viable, elongated smooth muscle cells from the swine carotid artery. Cells were prepared by papain digestion of pressurized arteries in calcium-free solution. After digestion, the arteries were everted, and fine strips were teased from the intimal surface of the media in calcium-free solution, releasing single cells. Viability was assessed by exclusion of trypan blue and by appearance under phase-contrast microscopy. By these criteria, approximately 20% of the isolated cells were viable. The most distinguishing and unexpected characteristic of these cells was their length. Mean length of the relaxed viable cells was 240.4 +/- 47.4 microns (SD, n = 76), which is much longer than previously reported for arterial smooth muscle cells. Calcium (1.6 mM) caused most of the viable cells to contract slightly, and the mean cell length in calcium was 194.4 +/- 57.7 microns. Cells in 1.6 mM calcium contracted substantially in response to 10 microM histamine or the calcium ionophore A23187 (10 microM), demonstrating that histamine receptors and the contractile apparatus were still functional.

scholarly journals Interleukin-6 and Leukemia Inhibitory Factor Induction of JunB Is Regulated by Distinct Cell Type-specific Cis-acting Elements

1999 ◽  
Vol 274 (40) ◽  
pp. 28697-28707 ◽  
Author(s):  
Robert M. Tjin Tham Sjin ◽  
Kenneth A. Lord ◽  
Abbas Abdollahi ◽  
Barbara Hoffman ◽  
Dan A. Liebermann
Keyword(s):  
Interleukin 6 ◽  
Leukemia Inhibitory Factor ◽  
Inhibitory Factor ◽  
Cell Type ◽  
Distinct Cell Type ◽  
Cis Acting ◽  
Distinct Cell ◽  
Cell Type Specific

Bombesin-like immunoreactivity in the avian gut and its localisation to a distinct cell type

1979 ◽  
Vol 61 (2) ◽  
pp. 213-221 ◽  
Author(s):  
C. M. Timson ◽  
J. M. Polak ◽  
J. Wharton ◽  
M. A. Ghatei ◽  
S. R. Bloom ◽  
...  
Keyword(s):  
Cell Type ◽  
Distinct Cell Type ◽  
Distinct Cell

scholarly journals A probabilistic gene expression barcode for annotation of cell-types from single cell RNA-seq data

2020 ◽  
Author(s):  
Isabella N. Grabski ◽  
Rafael A. Irizarry
Keyword(s):  
Gene Expression ◽  
Single Cell ◽  
Latent Variable ◽  
Cell Types ◽  
Marker Genes ◽  
Cell Type ◽  
Variable Model ◽  
Distinct Cell Type ◽  
Distinct Cell ◽  
Public Datasets

AbstractSingle-cell RNA sequencing (scRNA-seq) quantifies gene expression for individual cells in a sample, which allows distinct cell-type populations to be identified and characterized. An important step in many scRNA-seq analysis pipelines is the annotation of cells into known cell-types. While this can be achieved using experimental techniques, such as fluorescence-activated cell sorting, these approaches are impractical for large numbers of cells. This motivates the development of data-driven cell-type annotation methods. We find limitations with current approaches due to the reliance on known marker genes or from overfitting because of systematic differences between studies or batch effects. Here, we present a statistical approach that leverages public datasets to combine information across thousands of genes, uses a latent variable model to define cell-type-specific barcodes and account for batch effect variation, and probabilistically annotates cell-type identity. The barcoding approach also provides a new way to discover marker genes. Using a range of datasets, including those generated to represent imperfect real-world reference data, we demonstrate that our approach substantially outperforms current reference-based methods, in particular when predicting across studies. Our approach also demonstrates that current approaches based on unsupervised clustering lead to false discoveries related to novel cell-types.

scholarly journals Initial Cell Type Choice in Dictyostelium

2010 ◽  
Vol 10 (2) ◽  
pp. 150-155 ◽  
Author(s):  
Wonhee Jang ◽  
Richard H. Gomer
Keyword(s):  
Single Cells ◽  
Cell Types ◽  
Break Symmetry ◽  
Stalk Cell ◽  
Cycle Phase ◽  
Cell Type ◽  
Content Type ◽  
Initial Cell ◽  
Distinct Cell ◽  
A Cell

ABSTRACT Much remains to be understood about how a group of cells break symmetry and differentiate into distinct cell types. The simple eukaryote Dictyostelium discoideum is an excellent model system for studying questions such as cell type differentiation. Dictyostelium cells grow as single cells. When the cells starve, they aggregate to develop into a multicellular structure with only two main cell types: spore and stalk. There has been a longstanding controversy as to how a cell makes the initial choice of becoming a spore or stalk cell. In this review, we describe how the controversy arose and how a consensus developed around a model in which initial cell type choice in Dictyostelium is dependent on the cell cycle phase that a cell happens to be in at the time that it starves.

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