scholarly journals Separation of Spermatogenic Cell Types Using STA-PUT Velocity Sedimentation

10.3791/50648 ◽  
2013 ◽  
Author(s):  
Jessica M Bryant ◽  
Mirella L Meyer-Ficca ◽  
Vanessa M Dang ◽  
Shelley L Berger ◽  
Ralph G Meyer
Keyword(s):  
Cell Types ◽  
Spermatogenic Cell ◽  
Velocity Sedimentation

Testis Biopsy Image Analysis Effectively Quantifies Spermatogenic Cell Types

1997 ◽  
pp. 147-150 ◽  
Author(s):  
Edward D. Kim ◽  
William W. Lin ◽  
Jacki Abrams ◽  
Larry I. Lipshultz
Keyword(s):  
Image Analysis ◽  
Cell Types ◽  
Spermatogenic Cell ◽  
Biopsy Image

scholarly journals Isolation and separation of highly enriched fractions of viable mouse gastric parietal cells by velocity sedimentation.

1975 ◽  
Vol 65 (2) ◽  
pp. 428-438 ◽  
Author(s):  
L J Romrell ◽  
M R Coppe ◽  
D R Munro ◽  
S Ito
Keyword(s):  
Structural Integrity ◽  
Light And Electron Microscopy ◽  
Cell Types ◽  
Parietal Cells ◽  
Velocity Sedimentation ◽  
Isolated Cells ◽  
Adult Mice ◽  
Tissue Dissociation ◽  
Cell Fractions ◽  
Gastric Parietal Cells

Methods of tissue dissociation and cell separation have been modified to obtain highly enriched fractions of mouse gastric parietal cells. Suspension of gastric mucosal cells are prepared by pronase digestion of the glandular portion of the stomach from adult mice. By utilizing the velocity sedimentation technique to separate cells of different sizes it is possible to recovery parietal cells, which are larger than the other cell types, in fractions with purity of 75-95%. The homogeneity of cell fractions has been assessed by light and electron microscopy. The ability of the isolated cells to exclude the dye trypan blue, to incorporate labeled substrate, to consume oxygen, and to retain their structural integrity indicates that they are viable and still capable of functional activity.

scholarly journals Separation of helper and suppressor T lymphocytes on a ficoll velocity sedimentation gradient.

1976 ◽  
Vol 143 (5) ◽  
pp. 1199-1210 ◽  
Author(s):  
H Tse ◽  
R W Dutton
Keyword(s):  
T Cells ◽  
Cell Types ◽  
Velocity Sedimentation ◽  
Constant Number ◽  
Spleen Cells ◽  
Suppressor T Cells ◽  
Con A ◽  
Normal Spleen ◽  
Suppressor T Lymphocytes

A 5-20% Ficoll velocity sedimentation gradient has been successfully applied to separate concanavalin A (Con A)-induced helper; and suppressor T cells. When titrated into a constant number of fresh normal spleen cells responding to sheep erythrocytes, cells from the top pool show stimulatory effects while those from the bottom pool show inhibitory activity. Both activities are found to be Con A dependent and anti-theta sensitive. We conclude that Con A-induced helper and suppressor T cells are distinct subpopulations and such separation will allow further characterization of these cell types.

scholarly journals Isolation of Reed-Sternberg cells from lymph nodes of Hodgkin's disease patients

Blood ◽  
1989 ◽  
Vol 73 (1) ◽  
pp. 222-229
Author(s):  
G Sitar ◽  
E Brusamolino ◽  
C Bernasconi ◽  
E Ascari
Keyword(s):  
Lymph Nodes ◽  
Hodgkin's Disease ◽  
Mononuclear Cells ◽  
Density Gradient Centrifugation ◽  
Hodgkin’S Disease ◽  
Gradient Centrifugation ◽  
Cell Types ◽  
Velocity Sedimentation ◽  
Single Cell Suspension ◽  
Final Yield

This study describes a simple and relatively rapid method of purifying Reed-Sternberg (R-S) cells and their morphologic variants from the lymph nodes of patients affected by Hodgkin's disease. Our initial studies defined the optimal procedure for a quantitative disaggregation of Hodgkin's lymph nodes and the densities of R-S cells in several donors. These preliminary steps were helpful in the development of strategies for selectively concentrating R-S cells by density gradient centrifugation. We layered a single-cell suspension over Percoll of appropriate density, centrifuged these samples for 15 minutes, and collected a fraction enriched in R-S cells. Most of the R-S cells were distributed between densities of 1.060 and 1.072, with a peak at approximately 1.066 g/mL. R-S cells are denser than many mononuclear cells present in the lymph nodes of Hodgkin's patients and lighter than reactive cells such as eosinophils, mast cells, and neutrophils. However, the ranges of densities of these cell types overlap, making purification of R-S cells by isopyknic centrifugation impossible. Nevertheless, when this enriched fraction is further processed by velocity sedimentation in order to take advantage of the larger size of R-S cells as compared with all other cells, a substantial purification is achieved. We used three different velocity-sedimentation chambers to find the optimal conditions for obtaining the highest purity with a high final yield. The cells isolated by this method are viable, appear to be morphologically normal, and have been further characterized biologically.

Testis Biopsy Image Analysis Effectively Quantifies Spermatogenic Cell Types

1997 ◽  
Vol 157 (1) ◽  
pp. 147-150 ◽  
Author(s):  
Edward D. Kim ◽  
William W. Lin ◽  
Jacki Abrams ◽  
Larry I. Lipshultz
Keyword(s):  
Image Analysis ◽  
Cell Types ◽  
Spermatogenic Cell ◽  
Biopsy Image

scholarly journals Zinc Transport Differs in Rat Spermatogenic Cell Types and Is Affected by Treatment with Cyclophosphamide

2016 ◽  
Vol 95 (1) ◽  
pp. 22-22 ◽  
Author(s):  
A. M. Downey ◽  
B. F. Hales ◽  
B. Robaire
Keyword(s):  
Cell Types ◽  
Spermatogenic Cell ◽  
Zinc Transport

scholarly journals Isolation of Reed-Sternberg cells from lymph nodes of Hodgkin's disease patients

Blood ◽  
1989 ◽  
Vol 73 (1) ◽  
pp. 222-229 ◽  
Author(s):  
G Sitar ◽  
E Brusamolino ◽  
C Bernasconi ◽  
E Ascari
Keyword(s):  
Lymph Nodes ◽  
Hodgkin's Disease ◽  
Mononuclear Cells ◽  
Density Gradient Centrifugation ◽  
Hodgkin’S Disease ◽  
Gradient Centrifugation ◽  
Cell Types ◽  
Velocity Sedimentation ◽  
Single Cell Suspension ◽  
Final Yield

Abstract This study describes a simple and relatively rapid method of purifying Reed-Sternberg (R-S) cells and their morphologic variants from the lymph nodes of patients affected by Hodgkin's disease. Our initial studies defined the optimal procedure for a quantitative disaggregation of Hodgkin's lymph nodes and the densities of R-S cells in several donors. These preliminary steps were helpful in the development of strategies for selectively concentrating R-S cells by density gradient centrifugation. We layered a single-cell suspension over Percoll of appropriate density, centrifuged these samples for 15 minutes, and collected a fraction enriched in R-S cells. Most of the R-S cells were distributed between densities of 1.060 and 1.072, with a peak at approximately 1.066 g/mL. R-S cells are denser than many mononuclear cells present in the lymph nodes of Hodgkin's patients and lighter than reactive cells such as eosinophils, mast cells, and neutrophils. However, the ranges of densities of these cell types overlap, making purification of R-S cells by isopyknic centrifugation impossible. Nevertheless, when this enriched fraction is further processed by velocity sedimentation in order to take advantage of the larger size of R-S cells as compared with all other cells, a substantial purification is achieved. We used three different velocity-sedimentation chambers to find the optimal conditions for obtaining the highest purity with a high final yield. The cells isolated by this method are viable, appear to be morphologically normal, and have been further characterized biologically.

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