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Our experience is limited to the isolation and short-term culture of hepatocytes, and towards this end, our goal is to simply provide the information needed for the isolation of healthy, fully functional cells. Instructions for the performance of select functional assays will be added in the near future, as these will serve as benchmarks to assess the quality and consistency of isolated hepatocytes. We have successfully and consistently performed a number of assays in isolated hepatocytes, including:.

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Our experience is limited to the isolation and short-term culture of hepatocytes, and towards this end, our goal is to simply provide the information needed for the isolation of healthy, fully functional cells. Instructions for the performance of select functional assays will be added in the near future, as these will serve as benchmarks to assess the quality and consistency of isolated hepatocytes. We have successfully and consistently performed a number of assays in isolated hepatocytes, including:.
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Home | mouselivercells.com Reviews

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Our experience is limited to the isolation and short-term culture of hepatocytes, and towards this end, our goal is to simply provide the information needed for the isolation of healthy, fully functional cells. Instructions for the performance of select functional assays will be added in the near future, as these will serve as benchmarks to assess the quality and consistency of isolated hepatocytes. We have successfully and consistently performed a number of assays in isolated hepatocytes, including:.

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1

Video

http://www.mouselivercells.com/video.html

These videos demonstrate the cannulation, wash (HBSS), digestion (collagenase/DMEM), excision, and purification steps described in the associated protocol. All procedures were performed in an IACUC-sanctioned lab and in complete accordance with IACUC regulations. Please click on the video thumbnails to navigate to the respective video page. If you are using Internet Explorer 8 or earlier, once you click on a video link, please double-click on the actual video to begin playing, and click once to pause.

2

FAQs

http://www.mouselivercells.com/troubleshooting.html

1 Liver does not completely blanch/only one lobe blanches/odd blanching (vascular appearance)/other blanching anomalies. 2 Liver immediately swells (excessively) upon increasing flow rate after initial cannulation, even after IVC has been cut. Check the osmolality of your HBSS, and make sure you are actually perfusing HBSS, and not H20. Make sure the blood has not clotted already- is there any liquid coming out of the IVC? Make sure you did not introduce air into the system after cannulation- did you com...

3

Notes

http://www.mouselivercells.com/notes.html

1] Disposable, sterilized cell strainers:BD #352350 70um. Reusable cell strainers: Dual Mfg #US3-200S 74um. Brass frame/stainless steel mesh; #200 mesh/74um; 3" diameter x 1" depth). Disposable BD strainers fit over standard 50mL conical tubes. Reusable Dual Mfg strainers work best with 95mm diameter culture plates; the lip on the strainer frame sits on top of the 95mm plate perfectly, while 100mm plates are a bit too wide, causing the strainer to sit entirely on the bottom of the plate. 5] See formulati...

4

otherprotocols

http://www.mouselivercells.com/otherprotocols.html

Primary hepatocytes are highly versatile due to their high metabolic activity and diverse functions in vivo; many, if not all of these functions may be reproduced in vitro. Please pay close attention to the date stamps on these protocols. Updated versions, when provided, will have more recent time stamps. This page will be updated in the near future when I have polished up my remaining protocols and spreadsheet tools. Glycogen and gluconeogenesis protocols. Digestion of glycogen in primary hepatocytes.

5

Procedure

http://www.mouselivercells.com/procedure.html

Download the PDF version of the entire protocol here. PDF version of this protocol updated 2010-11-21; uploaded 2012-04-07. Minor differences in Troubleshooting section compared to website. Prepare all necessary reagents. For each mouse, you will need (see details for reagents in materials):. 60-70mL HBSS for initial flushing/washing; 37C. 90mL DMEM-low glucose (digestion medium); 37C. 120mL isolation medium; 4C. Again, note that these quantities are based upon the C57 strain of mice weighing 20-40g.

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Our experience is limited to the isolation and short-term culture of hepatocytes, and towards this end, our goal is to simply provide the information needed for the isolation of healthy, fully functional cells. Instructions for the performance of select functional assays will be added in the near future, as these will serve as benchmarks to assess the quality and consistency of isolated hepatocytes. We have successfully and consistently performed a number of assays in isolated hepatocytes, including:.

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