Preparation of gastrointestinal hormone-releasing hybridoma

我们尝试制造小肠干细胞和肠内分泌细胞的杂交瘤。首先，以IEC-6细胞系为材料，选择培养基，制备小肠干细胞所需噻咪啶的突变体。获得的突变体生长需要硫胺，如果没有硫胺，突变体在1周内死亡。然后，从大鼠小肠中纯化肠内分泌细胞。刮取大鼠小肠上皮细胞层，进行酶解。然后将分散的细胞以40%的倍率进行密度梯度超离心。获得了含有高浓度CCK生成细胞的部分，但该部分仍有污染细胞。现在我们尝试用细胞分选器从这个部分进一步纯化产生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.