Most traditional methods published for isolating hepatocytes use crude and partially purified enzyme preparations including various types of collagenase and other proteases. More recently the use of better characterized preparations of collagenase such as Types 1 and 4 have provided better results. All crude collagenase preparations can contain lot-variable contaminating proteases, esterases and other enzymes requiring researchers to pre-screen several lots of enzyme and/or continually modify isolation parameters and protocols.
The Hepatocyte Isolation System has been developed to provide researchers with a reliable, convenient, and consistent hepatocyte cell isolation system. By using the pre-optimized combination of enzymes contained in this kit, it is possible to minimize the lot-to-lot variation and improve the quality of the isolated hepatocytes. In addition, each lot is tested by isolating hepatocytes from adult rat to assure performance, reliability, and consistent yield of viable cells.
The method is based on that described by Berry, M.N., modified by Seglen, P.O. (Methods in Cell Biology, vol XIII, David M. Prescott ed., Academic Press, 1976; Chapter 4, "Preparation of Isolated Rat Liver Cells", pp 29-83), and further optimized in conjunction with several researchers.
This kit contains sufficient materials for five separate adult rat liver perfusions. For larger or smaller tissue applications, prepare proportionate volumes of reagents at each step and combine them in the same ratio as described in the protocol.
Storage and Stability:Store all components at 4°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
