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> Hepatocyte Isolation System (Tissue Dissociation/Cell Isolation), BioAssay™ Kit

Hepatocyte Isolation System (Tissue Dissociation/Cell Isolation), BioAssay™ Kit

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Référence H2006-02-1Kit

Conditionnement : 1Kit

Marque : US Biological

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H2006-02 Hepatocyte Isolation System (Tissue Dissociation/Cell Isolation), BioAssay™ Kit

Shipping Temp
Blue Ice
Storage Temp
4°C

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.

Kit Components:
H2006-02A: CMF-HBSS, 10X, 1x500ml
H2006-02B: Collagenase-Elastase Enzyme, 5x1vial
H2006-02C: DNAse 1, 1000U 5x1vial
H2006-02D: 0.15M MOPS, 1x75ml
H2006-02E: 7.5% Sodium Bicarbonate, 1x100ml
H2006-20F: Leibovitz L-15 (Powder), 1x1L

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.

Applications
Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
References
1. Alpini et al., "Recent Advances in the Isolation of Liver Cells", Hepatology (1994) 20:494-514. 2. Berry, M.N., Edwards, A.M., and Barritt, G.J.; Isolated Hepatocytes: Preparation, Properties and Applications, RH Burdon and PH Van Knippenberg, eds., Elsevier,Amsterdam, New York, Oxford, Chapt. 2, 1991. 3. Chen H-L, Wu H-l, Fon C-C, Chen P-J, Lai, M-Y, Chen D-S(1998) Long-term culture of hepatocytes from human adults. Biomedical Science 5:435-440. 4. DeRobertis, E.D.P., Saez, F.A. and DeRobertis, E.M.F.:Cell Biology, 6th ed., W.B. Saunders Co., Philadelphia, PA,1975.F5. reshney, R. Ian: Culture of Animal Cells, Alan R. Liss,Inc., New York, 2000.. 5. Hanks, J.H., and Wallace, R.E.: Proc. Soc. Exp. Biol. Med.,71, 196 1949.. 6. Jakoby, W.B., and Pastan, I.H.: Methods in Enzymology Vol. LVIII p. 121, Academic Press, 1979. 7. Le Cluyse EL (2001) Human hepatocyte systems for the in vitro evaluation of cytochrome P450 enzyme expression and regulation. Eur. J Pharm Sci 13:343-368. 8. Le Cluyse EL, PL Bullock, A Parkinson (1996) Strategies for restoration and maintenance of normal hepatic structure and function in long-term cultures of rat hepatocytes. Adv.Drug Del Rev 22:133-186. 9. Leibovitz, A.: The Growth and Maintenance of Tissue/Cell Cultures in Free Gas Exchange with the Atmosphere, Am.J. Hyg., 78, 173, 1963. 10. Seglen, P.O., Methods in Cell Biology, Vol. XIII, David M.Prescott ed., Ch. 4, pp. 29-83, Academic Press, 1976.