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Immortalized canine adipic mesenchymal stem cell line and constructing method thereof

A kind of adipose-derived mesenchymal stem cell and construction method technology, applied in the field of immortalized canine adipose-derived mesenchymal stem cell line and its construction, which can solve the problems of poor viability and difficult survival of canine adipose-derived mesenchymal stem cells, and achieve a shortened cell incubation period and good viability. , the effect of a large number of cells

Inactive Publication Date: 2016-07-20
NORTHWEST A & F UNIV
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Problems solved by technology

[0005] In order to solve the problem that the isolated canine adipose-derived mesenchymal stem cells have poor vigor and difficulty in survival as the number of passages increases, the purpose of the present invention is to provide an immortalized canine adipose-derived mesenchymal stem cell line and its construction method

Method used

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  • Immortalized canine adipic mesenchymal stem cell line and constructing method thereof
  • Immortalized canine adipic mesenchymal stem cell line and constructing method thereof
  • Immortalized canine adipic mesenchymal stem cell line and constructing method thereof

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Embodiment Construction

[0033]The invention provides an immortalized canine adipose-derived mesenchymal stem cell line, which uses canine adipose-derived mesenchymal stem cells as host cells, transfects pLOX-Ttag-iresTK lentiviral vector, and expresses SV40 large T antigen and obtains through continuous passage. Transformants with multilineage 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 construction of the immortalized canine adipose-derived mesenchymal stem cell line, cell morphological characteristics, gene and immune identification of markers, and differentiation induction test are described in detail below, which are explanations rather than limitations of the present invention.

[0034] 1. Construction of pLOX-Ttag-iresTK lentiviral vector

[0035] The pLOX-Ttag-iresTK lentiviral vector was purchased from Addgene.

[0036...

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Abstract

The invention discloses an immortalized canine adipic mesenchymal stem cell line and a constructing method thereof. The immortalized canine adipic mesenchymal stem cell line capable of expressing SV40 large T antigen genes and having multidirectional differentiation is obtained by: transfecting a lentiviral vector carrying SV40 large T antigen (Ttag) by using canine adipic mesenchymal stem cells as host cells, integrating the Tag genes into an adipic mesenchymal stem cell genome and carrying out continuous passage and screening. This cell line is still active in case of 50 in-vitro passages, is high in differentiation and proliferation performance, never ages or dies, can also save medium cost and is an immortalized cell line.

Description

technical field [0001] The invention belongs to the technical field of cell engineering, and in particular relates to an immortalized canine adipose-derived mesenchymal stem cell line and a construction method thereof. Background technique [0002] Pets play an irreplaceable emotional adjustment role in many modern families. However, many serious pet traumas, metabolic diseases and other problems have seriously affected the quality of life of pets. The current veterinary clinical treatment methods still cannot fundamentally solve the high incidence of internal and surgical diseases in many pets. Therefore, it is hoped that stem cell replacement therapy can solve these problems. [0003] Adipose-derived mesenchymal stem cells (Adipose-derived mesenchymal stem cells) are a type of adult stem cells existing in adipose tissue, capable of self-renewal, and can differentiate into osteoblasts, adipocytes, islet cells, etc. The characteristic is that allogeneic or even xenotransp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/867C12R1/91
CPCC12N5/0667C12N15/867
Inventor 华进联方佳魏于栋赵善廷
Owner NORTHWEST A & F UNIV
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