Preparation of gastrointestinal hormone-releasing hybridoma

我们试图制作一个由小肠干细胞和肠道内分泌细胞组成的杂交瘤。首先，以IEC-6细胞系为材料，通过筛选培养基制备出生长所需的硫胺化小肠干细胞突变体。获得的突变株生长需要硫胺，没有硫胺，它们在1周内死亡。然后从大鼠小肠中纯化肠内分泌细胞。取大鼠小肠上皮细胞层，酶消化。然后将分散的细胞应用于密度梯度超速离心法，每次40%。获得了一个含有高浓度产CCK细胞的组分，但该组分仍有污染细胞。现在，我们试图通过细胞分选机进一步从这一组分中纯化产生CCK的细胞。

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.