Preparation of gastrointestinal hormone-releasing hybridoma

我们试图制造一个小的生殖干细胞和肠内分泌细胞的杂交瘤。首先，用IEC-6细胞系和筛选培养基制备了小肠干细胞突变株。获得的突变体需要噻咪啶才能生长，没有噻咪啶，突变体在1周内死亡。刮取大鼠小肠上皮细胞层，酶消化。然后将分散的细胞施加在具有40%percall的密度梯度超离心上。获得了含有高浓度CCK产生细胞的级分，但该级分仍具有污染细胞。现在，我们试图进一步纯化CCK生产细胞从这一部分通过细胞分选仪。

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.