Establishment of a new method to induce the differentiation of embryonic pancreatic cells into mature endocrine cells.
Zhongguo yi xue ke xue yuan xue bao. Acta Academiae Medicinae Sinicae(2012)
摘要
OBJECTIVE:To establish a new culture method to induce the differentiation of embryonic pancreatic cells into mature endocrine cells.
METHODS:Mouse embryos at day 12.5 were used and embryonic pancreata were isolated. The isolated embryonic pancreata were cultured on the filter for 7 days, which floated in the dish containing medium. During culture, the expression of pancreas duodenum homeobox-1 (PDX-1), a pancreatic stem cell marker, was examined at day 1. The expression of neurogenin 3 (Ngn3), a pancreatic progenitor cell marker, was examined at day 3. The expressions of endocrine and exocrine markers, insulin, glucagon, and carboxypeptidase (CPA) were examined at day 7 by immunohistochemistry. The kinetics of pancreatic marker expression during culture was assayed by real-time PCR.
RESULTS:Many pancreatic stem cells still existed in embryonic pancreata cultured for 1 day; meanwhile, these pancreatic stem cells proliferated in high rate. A large amount of pancreatic progenitor cells were found in embryonic pancreata cultured for 3 days.Pancreatic stem/progenitor cells differentiated into mature endocrine and exocrine cells in embryonic pancreata after having been cultured for 7 days. Furthermore, the expression pattern of pancreatic marker is consistent with that in vivo.
CONCLUSION:We successfully established a new culture method, with which embryonic pancreatic cells can efficiently differentiate into mature endocrine cell.
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关键词
Insulin,Neurogenen 3,Pancreas,Pancreas duodenum homeobox-1
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