Preparation of gastrointestinal hormone-releasing hybridoma

1993 Fiscal Year Final Research Report Summary

• Project/Area Number: 04660086
• Research Category: Grant-in-Aid for General Scientific Research (C)
• Allocation Type: Single-year Grants
• Research Field: 応用生物化学・栄養化学
• Research Institution: KYOTO UNIVERSITY
• Principal Investigator: FUSHIKI Tohru Food Science & Tech, Kyoto Univ., Associate professor, 農学部, 助教授 (20135544)
• Co-Investigator(Kenkyū-buntansha): INOUE Kazuo Food Sci.& Tech, Kyoto Univ., Research associate, 農学部, 助手 (80213148) ; KAWADA Teruo Food Sci.& Tech, Kyoto Univ., Research associate, 農学部, 助手 (10177701)
• Project Period (FY): 1992 – 1993
• Keywords: Gastro intestinal hormone / hybridoma / small intestinal stem cell / CCK

Research Abstract

We tried to make a hybridoma of small intestineal stem cells and enteroendocrine cells. First, mutants of the small intestinal stem cells needed thimidine for growth were prepared with IEC-6 cell line and selecting culture medium. Obtained mutants were needed the thimidine for growth and without thimidine, they died within 1 week.
Then, enteroendocrine cells were purified from the rat small intestine. Epithelial cell layr of the rat small intestine were scraped and enzymatically digested. Dispersed cells were then applied on dencity gradient ultracentrifugation with 40% percall. A fraction containing high concentration of CCK producing cells was obtained but this fraction still has contaminant cells. Now we tried to further purify the CCK producing cells from this fraction by means of cell sorter.