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Method for large-scale culture of immortalized porcine hepatocyte

A large-scale culture and immortalization technology, applied in the field of medical biology

Inactive Publication Date: 2009-03-25
ZHEJIANG UNIV
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Problems solved by technology

[0005] However, in order to truly realize the application of immortalized porcine hepatocytes to replace primary porcine hepatocytes in bioartificial liver or hepatocyte transplantation, it is necessary to obtain a sufficient number of immortalized porcine hepatocytes with high activity and good functions, and Solving the immune rejection of xenogeneic hepatocytes is the key to the success of bioartificial liver or hepatocyte transplantation; therefore, in order to solve the above two problems, it is necessary to establish a method for large-scale cultivation of immortalized pig hepatocytes

Method used

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Embodiment Construction

[0017] Below in conjunction with specific implementation method the present invention is described in further detail:

[0018] Immortalized porcine hepatocytes were obtained as described in the literature (document: Pan Xiaoping, Du Weibo, Hao Shaorui, etc. Establishment and identification of immortalized porcine hepatocytes. Chinese Journal of Infectious Diseases, 2008, 26(7): 406- 409).

[0019] 1. Roller bottle culture of immortalized porcine hepatocytes, cell counting and cell viability analysis

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Abstract

The invention discloses a large-scale culture method of immortalized pork liver cells, which comprises: the immortalized pork liver cells collected are inoculated in a spinner bottle, rotating culture is carried out in a three-layer culture system, sodium alginate solution with a concentration of 1.5 percent to 2.0 percent is used for adjusting the concentration of the immortalized pork liver cells to 1 to 2.5 mulpied by 10<6> / mL; the immortalized pork liver cells obtained through readjustment of the concentration is prepared into immortalized pork liver cell micro-capsule by one-step method with alginic acid-chitosan; and the invention further discloses a microencapsulized immortalized pork liver cell. The microencapsulized immortalized pork liver cell can be used as liver cell source for the transplantation of biotype artificial liver or liver cell. The HCMV gBn1 antibody prepared has the advantages of strong specificity and high sensitivity.

Description

technical field [0001] The invention belongs to the field of medical biotechnology and relates to a method for large-scale cultivation of immortalized pig hepatocytes. Background technique [0002] In recent years, hepatocyte-based bioartificial liver or hepatocyte transplantation has become the focus and focus of basic and clinical research on the treatment of liver failure; hepatocytes are the biological part of the bioartificial liver and play a central role; Human hepatocytes can be regarded as the most ideal cell material for biological artificial liver because of their homology and good cell metabolism. Cells as the cell source of the bioartificial liver is the biggest obstacle. [0003] Due to the similar size, structure and biological characteristics of pig hepatocytes and human hepatocytes, their metabolic and immune functions are more similar to those of human hepatocytes; therefore, the current basic and clinical research on bioartificial liver (Document: Qian Y...

Claims

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Application Information

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IPC IPC(8): C12N5/06A61L27/38A61F2/02C12N5/071
Inventor 李兰娟潘小平杜维波喻成波吕国良虞晓鹏章益民
Owner ZHEJIANG UNIV
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