PMID: 3759794Sep 1, 1986Paper

A method for isolating large numbers of viable disaggregated cells from various human tissues for cell culture establishment

In Vitro Cellular & Developmental Biology : Journal of the Tissue Culture Association
R E Gibson-D'AmbrosioS M D'Ambrosio

Abstract

A method is described for the isolation of large numbers of viable disaggregated cells from human tissues. This method combined the mechanical action of a Stomacher Model 80 Lab Blender, 0.1 mg/ml trypsin or 0.5 mg/ml collagenase, and 0.1 mM [ethylene bis(oxyethylenenitrolo)]-tetraacetic acid (EGTA). Tissue (0.2 to 1.0 g) obtained from human fetal intestine, kidney, liver, lung, and skin were separately minced into approximately 1-mm3 pieces. The pieces were placed in a sterile bag containing 60 ml of calcium- magnesium-free phosphate buffered saline, the appropriate enzyme (0.1 mg/ml trypsin or 0.5 mg/ml collagenase) plus 0.1 mM EGTA, and 0.1% methylcellulose. The bag was then placed into the blender and mixed at a low speed for 3 to 20 min at room temperature. After a single cell suspension was observed by phase contrast microscopy, 10 ml of bovine calf serum was added to the cell suspension to inactivate the proteolytic enzymes. At this time 130 ml of cold Hanks' balanced salts solution containing 5% bovine calf serum was added and the entire cell suspension passed through a tissue sieve (100 mesh, 140 micron) and the cells collected by centrifugation. These cells were then resuspended into the appropriate culture medium. In...Continue Reading

References

Jan 1, 1979·Methods in Enzymology·M M Bashor
Jul 1, 1979·Proceedings of the Society for Experimental Biology and Medicine·H AutrupI C Hsu
Aug 1, 1986·In Vitro Cellular & Developmental Biology : Journal of the Tissue Culture Association·J M Raber, S M D'Ambrosio
Jun 1, 1949·Proceedings of the Society for Experimental Biology and Medicine·J H HANKS, R E WALLACE

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Citations

Jun 1, 1987·Klinische Wochenschrift·M DietelA Niendorf
Apr 1, 1987·In Vitro Cellular & Developmental Biology : Journal of the Tissue Culture Association·R E Gibson-D'AmbrosioS M D'Ambrosio
Jul 1, 1996·Veterinary Immunology and Immunopathology·S KrakowkaR Leunk
Oct 1, 1989·Journal of Photochemistry and Photobiology. B, Biology·S E OgutB M Sutherland
Oct 18, 2001·Journal of the National Cancer Institute·J R KanwarG W Krissansen
Jul 1, 1997·In Vitro Cellular & Developmental Biology. Animal·H J BoxbergerM J Sessler
Nov 25, 2006·Experimental and Toxicologic Pathology : Official Journal of the Gesellschaft Für Toxikologische Pathologie·Stéphane PédeboscqJean Cambar
Sep 18, 2008·International Journal of Cancer. Journal International Du Cancer·Haiming DingSteven M D'Ambrosio
Sep 18, 2008·International Journal of Cancer. Journal International Du Cancer·Haiming DingSteven M D'Ambrosio
Jan 1, 1989·Teratogenesis, Carcinogenesis, and Mutagenesis·G WaniS M D'Ambrosio
Jul 1, 1992·In Vitro Cellular & Developmental Biology : Journal of the Tissue Culture Association·R E Gibson-D'Ambrosio, S M D'Ambrosio
Dec 10, 1998·Journal of Biomedical Science·H L ChenD S Chen

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