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Immortalized porcine pancreatic stem cell line and construction and differentiation methods thereof

A stem cell line, pig pancreas technology, applied in the field of cell engineering, can solve the problems of apoptosis, poor vitality, difficult to survive, etc., and achieve the effect of reducing blood sugar concentration, maintaining blood sugar level, and improving vitality

Active Publication Date: 2011-02-16
陕西九州细胞基因工程有限公司
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Problems solved by technology

However, as the number of passages increases, the isolated pancreatic stem cells become less and less active, and eventually undergo apoptosis, making it difficult to survive.

Method used

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  • Immortalized porcine pancreatic stem cell line and construction and differentiation methods thereof
  • Immortalized porcine pancreatic stem cell line and construction and differentiation methods thereof
  • Immortalized porcine pancreatic stem cell line and construction and differentiation methods thereof

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Embodiment Construction

[0037]The invention provides an immortalized porcine pancreatic stem cell line, which uses porcine pancreatic stem cells as host cells, transfects pCI-neo-hTERT eukaryotic expression vector, and expresses human telomerase reverse transcriptase obtained through G418 screening and has multiple transformants to differentiation potential. The cell line still has good vigor after more than 50 passages in vitro, maintains division and proliferation, does not undergo senescence and apoptosis, and is an immortalized cell line. The following is a detailed description of the construction of the immortalized porcine pancreatic stem cell line, cell morphological characteristics, gene and immune identification of markers, induction of differentiation, and blood sugar changes in the transplanted diabetes model, which is an explanation of the present invention rather than limited.

[0038] 1. pCI-neo-hTERT eukaryotic expression vector

[0039] pCI-neo-hTERT can be constructed by vector lig...

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Abstract

The invention discloses an immortalized porcine pancreatic stem cell line and construction and differentiation methods thereof. In the immortalized porcine pancreatic stem cell line, porcine pancreatic stem cells are taken as host cells and are transfected with a pCI-neo-hTERT eukaryotic expression vector, and the human telomerase reverse transcriptase screened by G418 is expressed and has a transformant with multi-directional differentiation potentiality. The cell line also has higher activity after transferring for over 50 generations in vitro, keeps split proliferation, does not generate aging or apoptosis, and is an immortalized cell line. The cell line can be differentiated to form functional islet cell mass after induction, can reduce the blood glucose concentration after being transplanted into a mouse model suffering from diabetes and can maintain a normal blood glucose level within two weeks.

Description

technical field [0001] The invention belongs to the technical field of cell engineering, in particular to an immortalized porcine pancreatic stem cell line and its construction and differentiation method. Background technique [0002] Diabetes is a global medical problem. At present, although there are many methods for the treatment of diabetes, none of them can fundamentally solve the problem. Organ transplantation is a relatively good solution, but it is mainly limited by the shortage of donors and immune rejection. Therefore, people hope that stem cell replacement therapy can solve the problems of donor shortage and immune rejection: stem cells have strong proliferative ability, sufficient quantity, and relatively low transplant rejection. [0003] Pancreatic stem cells (Pancreatic stem cells) are a type of adult stem cells that exist in fetal and adult pancreatic tissues. They can self-renew and have multi-directional differentiation potential. However, human pancreat...

Claims

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Application Information

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IPC IPC(8): C12R1/91C12N15/52C12N15/85C12N5/10
Inventor 曹晖楚元奎华进联窦忠英
Owner 陕西九州细胞基因工程有限公司
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