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Human foetus pancreatic stem cell line and its establishing method

A stem cell line and stem cell technology, applied in the field of human fetal pancreatic stem cell line and its establishment, can solve the problems of inconsistent identification results of pancreatic stem cells

Inactive Publication Date: 2005-05-04
NORTHWEST A & F UNIV
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Problems solved by technology

However, considering that nerve, muscle, mesenchymal and adult pancreatic satellite cells also express nestin, Andreas et al. believe that (Am J Physinol Metab. 2003, 284: E259-E266.) These cells are not unique to the pancreas and are derived from The nestin-positive cells of pancreatic islets may have the potential of transdifferentiation into pancreatic endocrine cells
[0003] To sum up, the mammalian pancreatic stem cells have not been fully characterized at present, the isolation, purification and expansion methods of pancreatic stem cells are all under exploration, and the identification results of pancreatic stem cells obtained are not consistent. Pancreatic stem cell line reports

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  • Human foetus pancreatic stem cell line and its establishing method
  • Human foetus pancreatic stem cell line and its establishing method
  • Human foetus pancreatic stem cell line and its establishing method

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Embodiment Construction

[0046] In order to understand the present invention more clearly, the present invention will be further described in detail below in conjunction with specific examples completed by the inventors according to the above technical solutions.

[0047] 1. Establishment of human fetal pancreatic stem cell lines

[0048] 1.1 Isolation and cloning of primary human fetal pancreatic stem cells

[0049] Aseptically collect pancreas tissue from normal aborted fetuses aged 4 to 6 months, wash it repeatedly 5 to 6 times with 0.1M PBS(-) containing double antibodies (penicillin 80,000 U / ml, 100,000 U / ml), and cut the tissue pieces until 1mm 3 . Digest with 0.1% type IV collagenase (collegenIV, sigma) for 15-40 min, and terminate the digestion when a large number of single cells are seen under the microscope. The digested suspension was filtered through a 0.2 mm pore size filter, and centrifuged (1000 r.p.m, 5 min). Discard the supernatant, dilute the cells in the centrifuge tube and inoc...

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Abstract

This invention opens one establishing method of one fetal pancreas stem cell lines. epithelioid stem cell of fetal pancreas comes from fetal pancreas organization in normal abortion, in vitro culture can be clonned infinitely, and possesses the potential of multi-directional differentiation; after testing and analysis of morphological character, growth characteristics, immunohistochemistry characteristics, RT-PCR character, directional induced differectiation characters, chromosome karyotypes, DNA array, it is proved that this fetal pancreas stem cell is human originated fetal pancreas stem cell; this invention can provide affulent seed cells for researches on mammalian pancreas development and diabetes origination and dibetes treatment by reproducing Langerhans' islands' micro organs. It also provides an efficient and reliable method for establishing pancreas stem cell lines of creodonts and human beings.

Description

technical field [0001] The invention belongs to the field of human and mammalian cell (tissue) engineering in the subject of biotechnology, and relates to in vitro separation, purification and rapid expansion of human and mammalian pancreatic stem cells, in particular to a human fetal pancreatic stem cell line and its establishment method. Background technique [0002] Diabetes mellitus (DM) is one of the major diseases that endanger human health, but traditional medicine and insulin replacement therapy cannot fundamentally cure the disease. In recent years, with the continuous improvement of islet transplantation technology, it has become possible to cure type I and some type II diabetes (Shapiro et al.N Engl J Med.2000, 27(4):230~238.), and islet donors The contradiction of shortage is also more prominent. Isolating and cloning human and mammalian pancreatic stem cells in vitro as seed cells and inducing them to differentiate into functional islets is an effective way to ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/39A61P3/10C12N5/073C12Q1/68
Inventor 效梅窦忠英安立龙
Owner NORTHWEST A & F UNIV
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