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Cultivation and Pro-maturation of Pancreatic Endocrine Progenitor Cell

Author: WangLei
Tutor: ZhaoYuPei
School: Peking Union Medical College , China
Course: Basic surgical
Keywords: Precursor cells Endocrine cells Pancreatic cells Medical Diabetes model Insulin secretion Dissertation Stem Cell Culture Gene transfection Renal subcapsular transplantation
CLC: R329
Type: PhD thesis
Year: 2004
Downloads: 104
Quote: 0
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Abstract


Background and AimDiabetes Mellitus(DM), a great menace to human beings, is a disease caused by the paucity and/or malfunction of pancreatic endocrine β cells. Cell replacement therapy plays an important role in the therapy of diabetes mellitus,though it face the problem of donor shortage. Pancreatic stem cell, with its reproductive ability, cast a light on the DM therapy. How to cultivate pancreatic stem cells conveniently and promote its maturation is the key point before its clinical usage. In this experiment, we choosed the pancrea of fetal SD rat and human bone marrow mesenchymal stem cell(BMSC) as the target of our research to find a path to solve the problems.MethodsWe selected the collagenase digestion method as the main way to cultivate pancreatic stem cells. To identify the cultivated cell line ,immunofluorescence,dithizone staining ,radioimmunoasssay,and electron microscope were performed.The cultivated cell line was transfected with recombined adenovirus plasmid , into which VEFG gene was incorporated.We observed the proliferation potential of cell line with MTT method. The cells with different kinds of cultivation were divided into four groups:UD,D,GFP,and VEGF.Then these 4 kinds of cells were implanted into the renal subcapsular space of BALB/c athymic mouse. The blood glucose(BG) , body weight(BW) ,survival period and insulin level were recorded and analyzed to determine the effect of

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