Isolation of rat Kupffer cells: a combined methodology for highly purified primary cultures
| Autor: | H. Roumpaki, Elias A. Kouroumalis, George Kolios, C. Xidakis, Vassilis Valatas |
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| Rok vydání: | 2003 |
| Předmět: |
Male
Kupffer Cells Cell Cell Culture Techniques Cell Separation Elutriation Biology Esterase Nitric oxide Rats Sprague-Dawley chemistry.chemical_compound Immunophenotyping Cell Adhesion medicine Animals Differential centrifugation HEPES Cell Biology General Medicine Rats Specific Pathogen-Free Organisms medicine.anatomical_structure Biochemistry chemistry Cell culture Plastics |
| Zdroj: | Cell Biology International. 27:67-73 |
| ISSN: | 1065-6995 |
| Popis: | We report a four-step procedure that optimizes the methodology for isolation of highly purified rat Kupffer cells (KC). We combined the previously reported techniques of enzymatic tissue treatment, density gradient centrifugation, centrifugal elutriation and selective adherence. ED-2 immunophenotyping and non-specific esterase histochemistry were used for cell identification. This combination resulted in a satisfactorily high yield of 80-100 x 10(6)KCs per liver, over 95% positive for ED-2 and 98% viable cells. Cultures of isolated KCs were functionally intact and exhibited a concentration and time-dependent LPS-induced TNF-alpha and nitric oxide production. |
| Databáze: | OpenAIRE |
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