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  • Title: Long-term explant culture of human colon and a 3-step transformation model for rat colonic epithelium.
    Author: Sakamoto K, Resau JH, Shamsuddin AM, Yuasa Y, Hoshino H, Nakano G, Nagamachi Y.
    Journal: Pathobiology; 1991; 59(6):404-11. PubMed ID: 1657027.
    Abstract:
    Our previous studies have suggested that colonic epithelium from rodents pretreated in vivo with suboptimal doses of carcinogen could be more easily maintained in explant culture. Transformation of colonic epithelium from these explants may be induced by subsequent exposure to additional genotropic agents. Therefore, we describe the development of a 3-step transformation model which uses (1) in vivo pretreatment with a suboptimal dose of 1,2-dimethylhydrazine (DMH) followed by (2) in vitro organ culture and exposure to xenotropic murine sarcoma virus (X-MSV), and finally (3) xenograft maintenance in nude mice to allow sufficient time for transformation. Male Wistar rats were injected intramuscularly with 20 mg DMH/kg 10 times per week followed by the removal and explant culture of the colon, which was then treated in vitro with X-MSV, and transplanted into nude mice after 1 week of culture. All the nude mice (n = 6) transplanted with rat colon explants contained viable xenograft explant epithelium and 1 of the 6 showed transformation. Our results demonstrate that the epithelium from animals pretreated with suboptimal doses of carcinogens can be easily transformed. We also demonstrate that human colonic epithelium is viable for an extended period of time in this model. Based on these results, we hypothesize that such a 3-step transformation model is applicable for carcinogenesis studies of various organs from different species, including human if one uses dysplastic or 'pre-neoplastic initiated' tissues obtained at surgery (e.g., ulcerative colitis; Barret's esophagus, etc.).
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