In vitro cultivation of human islets from expanded ductal tissue.
نویسندگان
چکیده
A major obstacle to successful islet transplantation for both type 1 and 2 diabetes is an inadequate supply of insulin-producing tissue. This need for transplantable human islets has stimulated efforts to expand existing pancreatic islets and/or grow new ones. To test the hypothesis that human adult duct tissue could be expanded and differentiated in vitro to form islet cells, digested pancreatic tissue that is normally discarded from eight human islet isolations was cultured under conditions that allowed expansion of the ductal cells as a monolayer whereupon the cells were overlaid with a thin layer of Matrigel. With this manipulation, the monolayer of epithelial cells formed three-dimensional structures of ductal cysts from which 50-to 150- micrometer diameter islet-like clusters of pancreatic endocrine cells budded. Over 3-4 weeks culture the insulin content per flask increased 10- to 15-fold as the DNA content increased up to 7-fold. The cultivated human islet buds were shown by immunofluorescence to consist of cytokeratin 19-positive duct cells and hormone-positive islet cells. Double staining of insulin and non-beta cell hormones in occasional cells indicated immature cells still in the process of differentiation. Insulin secretion studies were done over 24 h in culture. Compared with their basal secretion at 5 mM glucose, cysts/cultivated human islet buds exposed to stimulatory 20 mM glucose had a 2.3-fold increase in secreted insulin. Thus, duct tissue from human pancreas can be expanded in culture and then be directed to differentiate into glucose responsive islet tissue in vitro. This approach may provide a potential new source of pancreatic islet cells for transplantation.
منابع مشابه
Isolation and in vitro cultivation of human skin Keratinocytes and preparation of epidermal sheet
متن کامل
The application of discriminant analysis in differentiation of fibroadenoma and ductal carcinoma of breast tissue using ultrasound velocity measurement
Background: Ultrasound propagation velocity was measured experimentally in normal, fibroadenoma and ductal carcinoma breast tissues, in order to distinguish normal breast tissue from tumors. Materials and methods: In quantitative measurements of ultrasound velocity, 403 breast tissue images were selected, comprising 130 normal breast tissue, 130 fibroadenoma, and 143 ductal carcinoma tumors. Th...
متن کاملIsolation and Cultivation of Adult Human Keratinocyte Stem Cells for Regeneration of Epidermal Sheets
Background: Keratinocyte stem cell is one of the adult stem cells that inhabits the skin and contributes to skin function and renewal. Adult stem cells are best defined by their capacity to self-renew, and to maintain tissue function for a long period of time. These findings indicate the importance of these cells for clinical applications including regenerative medicine, tissue engineering and ...
متن کاملMONOPHASIC STATE OF INSULIN SECRETION IN ISOLATED RAT ISLETS OF LANGERHANS BY K+, Ca2+ AND GLUCOSE
Isolation of rat islets of Langerhans was carried out by using collagenase and the destruction of exocrine tissue was performed through the injection of Hanks' solution into the common bile duct. The identification of the islets was done by techniques including the use of a stereomicroscope against a black background. Vital staining was done by an injection of neutral red solution through t...
متن کاملO-19: Proliferation of Small Population of Spermatogonial Stem Cells in Azoospermic Patients
Background: With treatment success in young boys with cancer, long-term effects of cancer treatment have found importance in pediatric oncology. Temporary or permanent infertility after treatment is an important subject in childhood and adult cancer patients which decrease quality of life. The one approach to overcome infertility in these cases is to cryopreserve small biopsy testicular tissue ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Proceedings of the National Academy of Sciences of the United States of America
دوره 97 14 شماره
صفحات -
تاریخ انتشار 2000