Cell cloning of Barrett's esophagus stem cell, gastric cardia stem cells and normal esophagus stem cells. Homo sapiens

Barrett’s esophagus confers significant risk of esophageal adenocarcinoma. We have established the cloning of patient-matched stem cells of Barrett’s, gastric, and esophageal epithelium. Barrett's esophagus stem cells (BE), gastric cardia stem cells (GC) and normal esophagus stem cells (Eso) from 12 patients were cloned (For BE: 12 patients, GC: 12 patients and Eso: 2 patients). Keratin 5 positive and Keratin 7 positive cells were cloned from human fetal esophageal epithelium. Using air liquid interface culture system, stem cells were induced to differentiate into mature epithelial structures. Overall design: We cultured the cloned patient-matched stem cells of Barrett’s, gastric, esophageal epithelium, fetal esophagus cells (krt5 and krt7 + cells) and performed the ALI structure. Total RNA were isolated from them. We used Affymetrix chips (HuEx-1_0-st-v2) to hybridize 2 samples from each stem cell and ALI strucure.

巴雷特食管（Barrett’s esophagus）会显著增加食管腺癌（esophageal adenocarcinoma）的发病风险。本研究已成功构建巴雷特食管、胃及食管上皮的患者匹配型干细胞系。本研究从12例患者体内分离得到巴雷特食管干细胞（Barrett’s esophagus stem cells, BE）、胃贲门干细胞（gastric cardia stem cells, GC）与正常食管干细胞（normal esophagus stem cells, Eso）并完成克隆，其中BE组对应12例患者，GC组对应12例患者，Eso组对应2例患者；同时从人类胎儿食管上皮中克隆得到角蛋白5（Keratin 5, KRT5）阳性及角蛋白7（Keratin 7, KRT7）阳性细胞。借助气液界面培养系统（air liquid interface culture system），可诱导上述干细胞分化为成熟上皮结构。
实验整体设计如下：我们对构建的巴雷特食管、胃、食管上皮患者匹配型干细胞系，以及胎儿食管上皮来源的角蛋白5、7阳性细胞进行培养，并完成气液界面（air liquid interface, ALI）结构构建；随后从上述所有样本中提取总RNA。我们使用Affymetrix HuEx-1_0-st-v2芯片对每类干细胞及气液界面结构的2份样本进行杂交实验。