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KMID : 1204320090250030237
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Laboratory Animal Research
2009 Volume.25 No. 3 p.237 ~ p.242
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Primary Bovine Cell Cultures Using Various Digestion Conditions
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Jeon Kyong-Leek
Hwang Kyu-Kye
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Abstract
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We investigated various culture conditions to determine most appropriate method of culturing primary cells from bovine fetus. Organs were extracted from a fetus, minced and digested with several proteolytic enzymes (trypsin, collagenase and dispase). Removing of red blood cell (RBC) was carried out either by dilution method (dilution of minced tissues with culture medium) or by lysis method (treatment of minced tissues with a reagent containing NH₄Cl in HEPES). The dilution method was almost as efficient in removing RBC as the lysis method. But the lysis method damaged the cells greatly, resulting in very low survival rate for digested cells. The optimal digestive conditions differed slightly but were very similar for different organs. Trypsin digested tissues very quickly but greatly damaged cells. In the case of minced kidney tissues, treatment with 0.25% collagenase for 30 minutes followed by application of 0.01% dispase for 10 minutes resulted in most efficient digestion with minimal damage to digested cells.
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KEYWORD
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Bovine fetus, primary cell culture, collagenase, dispase, enzyme digestion
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