A simple non-enzymatic method for the isolation of high yields of functional rat hepatocytes |
| |
Authors: | Kravchenko L Petrenko A Shanina I Fuller B |
| |
Affiliation: | Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of the Ukraine, 23 Pereyaslavskaya St, Kharkov 310015, Ukraine. |
| |
Abstract: | A method for isolation of rat hepatocytes using liver perfusion by ethylenediamine tetra-acetic acid (EDTA)-containing sucrose solution and mechanical tissue disaggregation by controlled vibration (MVD) is described. The yields of hepatocytes produced by this method were similar to those obtained using collagenase perfusion. The cells had well preserved membrane integrity as judged by the trypan blue staining test (91 +/- 4%), ATP contents, rates of endogenous respiration and enzyme leakage that indicated they were functional cells. There was little evidence of expression of latent damage when the cells were stored either at 37 degrees C (by pre-incubation) or at 4 degrees C. This method can be used to isolate high yields of functional cells from rat liver if the collagenase perfusion technique is not available. |
| |
Keywords: | isolated rat hepatocytes mechanical disaggregation vibration sucrose buffer viability indexes |
本文献已被 PubMed 等数据库收录! |