| Monolayer cell culture of freshly isolated adipocytes using extracellular basement membrane components. | |
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MedLine Citation:
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PMID: 7616129 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Cell biological techniques requiring cells attached to surfaces, such as monolayer cell culture, microspectrofluorometry, and confocal microscopy, have not been readily available for use on adipocytes because they float and tend to lyse when attached to charged non-biological surfaces. A new method for attaching freshly isolated rodent adipocytes to thermanox plastic surfaces using Matrigel (a defined mixture of extracellular matrix components that resembles the basal lamina surrounding adipocytes in vivo) is described. The method takes advantage of an unusual physical characteristic of Matrigel, i.e., that it is a liquid at cold temperatures and a hydrated gel at higher temperatures. To attach the isolated cells, chilled thermanox plastic coverslips were coated with a thin uniform layer of ice-cold Matrigel and inverted into warm floating adipocytes. Adipocytes floated up against the liquid Matrigel and became immediately attached when the Matrigel changed to a gel in response to the warmth of the cells and media. Cell volume measurements of the attached versus freshly isolated cells indicate no significant difference in the centroid cell volume of the attached cells. This indicates that the method does not select for small or large cells. Adipocytes maintained for 6 days in culture did not display any change in their size or differentiated microscopic appearance. The relative concentrations of major proteins in silver-stained SDS-PAGE gels and several differentiation state-dependent proteins, including ATP-citrate lyase, carbonic anhydrase III (CA III), adipocyte lipid binding protein (ALBP), and pyruvate carboxylase, were examined. No significant change was observed in the relative concentrations of these proteins when the matrigel-cultured adipocytes were compared to freshly isolated cells.(ABSTRACT TRUNCATED AT 250 WORDS) |
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Authors:
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S A Hazen; W A Rowe; C J Lynch |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Journal of lipid research Volume: 36 ISSN: 0022-2275 ISO Abbreviation: J. Lipid Res. Publication Date: 1995 Apr |
Date Detail:
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Created Date: 1995-08-18 Completed Date: 1995-08-18 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 0376606 Medline TA: J Lipid Res Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 868-75 Citation Subset: IM |
Affiliation:
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Department of Cellular and Molecular Physiology, Milton S. Hershey Medical Center and College of Medicine, Pennsylvania State University, Hershey 17033, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Adipocytes
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cytology* Animals Basement Membrane Cell Adhesion Cells, Cultured Extracellular Matrix Rats Rats, Sprague-Dawley |
| Grant Support | |
ID/Acronym/Agency:
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DK02069/DK/NIDDK NIH HHS; DK43402/DK/NIDDK NIH HHS |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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