CAJ | 학술논문

背景:胚胎胰腺组织具有来源广泛,β细胞增殖分化能力强,低免疫排斥性等优点.目的:探索小鼠胚胎胰腺组织的分离技术,观察其对糖尿病模型小鼠的血糖调节作用.方法:体视显微镜下分离 E11.5～E16.5C57BL/6小鼠胰腺组织.链唑霉素诱导雄性C57BL/6小鼠建立糖尿病模型,随机分为2组:移植组模型小鼠肾被膜下移植5个E16.5胰腺组织,假手术组模型小鼠肾被膜下注入0.05mL RPMl1640培养液.移植组小鼠血糖水平≤11.2 mmol/L后,利用IPGTT和IPITT方法检测移植后胚胎胰腺组织的内分泌功能,并摘除移植物观察血糖变化.结果与结论:体视显微镜下可分离出较完整的E11.5～E16.5小鼠胰腺组织,≤E12.5 d小鼠胚胎胰腺组织的形态和颜色均难以与周围组织分辨,需根据其与毗邻脏器的关系仔细辨别:>E12.5 d的小鼠胚胎胰腺已初具形态,颜色略发白.组织学和ELISA分别显示胚胎胰腺组织可表达并分泌胰岛素,其表达强度随发育时间逐渐增加.E16.5小鼠胰腺组织移植能有效地控制受体的血糖水平,使受体的体质量和糖耐量恢复正常;胚胎胰腺在受体的肾被膜下可生长发育,摘除的移植物胰岛素和胰高血糖素的表达均较移植前增强.说明胚胎胰腺组织可能成为治疗糖尿病的种子来源.
배경:배태이선조직구유래원엄범,β세포증식분화능력강,저면역배척성등우점.목적:탐색소서배태이선조직적분리기술,관찰기대당뇨병모형소서적혈당조절작용.방법:체시현미경하분리 E11.5～E16.5C57BL/6소서이선조직.련서매소유도웅성C57BL/6소서건립당뇨병모형,수궤분위2조:이식조모형소서신피막하이식5개E16.5이선조직,가수술조모형소서신피막 하주입0.05mL RPMl1640배양액.이식조소서혈당수평≤11.2 mmol/L후,이용IPGTT화IPITT방법검측이식후배태이선조직적내분비공능,병적제이식물관찰혈당변화.결과여결론:체시현미경하가분리출교완정적E11.5～E16.5소서이선조직,≤E12.5 d소서배태이선조직적형태화안색균난이여주위조직분변,수근거기여비린장기적관계자세변별:>E12.5 d적소서배태이선이초구형태,안색략발백.조직학화ELISA분별현시배태이선조직가표체병분비이도소,기표체강도수발육시간축점증가.E16.5소서이선조직이식능유효지공제수체적혈당수평,사수체적체질량화당내량회복정상;배태이선재수체적신피막하가생장발육,적제적이식물이도소화이고혈당소적표체균교이식전증강.설명배태이선조직가능성위치료당뇨병적충자래원.
BACKGROUND: Embryonic pancreatic tissue is characterized by its abundance, potent in proliferation & differentiation, and minimal immunological rejection. It is widely considered as potential pancreatic endocrinological stem cells resource for treating diabetes mellitus.OBJECTIVE: To investigate the embryonic mouse pancreatic tissue isolation technique and observe the recipients' blood glucose regulatory effects of the grafted embryonic pancreas in an experimental diabetes mellitus mouse model.METHODS: Pancreatic tissue from C57B1/6 mouse embryos at embryonic days 11.5-16.5 was isolated under the stereomicroscope. C57BL/6 mouse models of streptozocin-induced diabetes mellitus were established and then randomly divided into two groups: transplantation group, in which, five pieces of pancreatic tissue of mice at embryonic 16.5 days were transplanted into mouse renal capsule, and sham-operated control group, in which, 0.05 mL RPMI1640 culture medium was injected into mouse renal capsule. When blood glucose level of the transplantation group mouse was≤ 11.2 mmol/L, the endocrine function of embryonic pancreatic tissue transplanted was detected by IPGTT and IPITT methods and then the transplanted graft was removed for observing the blood glucose relapse.RESULTS AND CONCLUSION: Nearly intact pancreatic tissue of mice at embryonic days 11.5-16.5 could be isolated through the use of stereomicroscope. Pancreatic tissue morphology and color of mice ≤ embryonic 12.5 days were difficultly distinguished from adjacent tissue and they could only be isolated carefully according to the relationship with adjacent organs. Pancreatic tissue of mice > embryonic 12.5 days exhibited initial endocrinological tissue morphology mimic white cauliflower. Histological and ELISA examinations showed that embryonic pancreatic tissue could express and secrete insulin and the insulin level was gradually increased with developmental time. Embryonic pancreatic tissue could grow beneath the recipient renal capsule. The insulin and glucagon expression in the post-transplantational pancreatic tissue graft was increased compared with prior to transplantation. These results suggest that pancreatic tissue is a potential stem cell resource for treating the diabetes mellitus.