Volume 14, Issue 2 (Scientific Journal of Hamadan University of Medical Sciences-Summer 2007)                   Avicenna J Clin Med 2007, 14(2): 16-22 | Back to browse issues page

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Shirvani Farsani Z, Mansouri K, Bidmeshkipour A, Mostafaie A. Isolation and Primary Culture of Rat Hepatocytes Using Kiwifruit Actinidin. Avicenna J Clin Med. 2007; 14 (2) :16-22
URL: http://sjh.umsha.ac.ir/article-1-423-en.html
Abstract:   (1468 Views)

Introduction & Objective: Isolation of cells from different tissues rely on proteolytic enzymes mainly collagenases that selectively digest collagen fibers of extra-cellular matrix. It is important to find new and proper collagenases from plant sources. In the present research actinidin, a cysteine protease abundant in Kiwifruit, was used to isolate and culture of rat hepatocytes.

Materials & Methods: Different concentrations of actinidin was used to isolate rat hepatocytes by one or two-step perfusion method. The viability of the separated cells was examined by the trypan blue test. The isolated rat hepatocytes were cultured on collagen coated plates in William´s E medium. The morphology of hepatocytes was examined microscopically after staining with the Papanicolaou method.

Results: Actinidin in the concentration of 0.4 mg/ml in two-step perfusion method properly isolated hepatocytes from rat liver. The viability of isolated hepatocytes that successfully cultured in collagen coated plates was 90-95 percent.

Conclusion: These results showed that actinidin is a proper protease for isolation of hepatocytes. In addition, purification of this enzyme is simpler than the collagenases.

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Type of Study: Original | Subject: Special

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