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Construction method of human lung teterocyte line

A construction method and cell line technology, which are applied in the field of human lung special network cell line construction, can solve the problems of time-consuming separation and purification in the extraction process, input a lot of manpower and material resources, and difficulty in ensuring the purity and stability of extracted cells, so as to reduce the pulmonary Inflammation, inhibition of apoptosis, and promotion of vascular endothelial cell proliferation

Pending Publication Date: 2022-03-25
ZHONGSHAN HOSPITAL FUDAN UNIV
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  • Description
  • Claims
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Problems solved by technology

[0002] The current research on the function of the special cells is to use the special cells extracted from different organs (lung, kidney, heart, etc.) of different species of animals (such as mice, rats, pigs, etc.), the extraction process is time-consuming and expensive every time Extraction requires separation, purification and identification, which requires a lot of manpower and material resources
Since multiple separations and extractions of primary cells are performed by different personnel, it is difficult to guarantee the purity and stability of the extracted cells

Method used

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  • Construction method of human lung teterocyte line
  • Construction method of human lung teterocyte line
  • Construction method of human lung teterocyte line

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Embodiment

[0050] The invention provides a method for constructing a human lung-specific cell line, comprising the following steps:

[0051] 1. The staged adherence method is adopted when extracting the primary special cells: take the discarded tissue after the operation, and cut it into 1mm in normal saline under sterile conditions 3 The tissue pieces were digested in 2mg / ml type II collagenase at 37°C for 30 minutes on a shaker, filtered through a 70-micron pore size filter, centrifuged at 1500rpm for 5 minutes, the supernatant was discarded, and the cell pellet was collected. 40ng / ml of EGF and / or FGF, 2mol / L of glutamine and 100mg / ml of penicillin / streptomycin in DMEM / F12 culture medium for 30 minutes, after the fibroblasts adhere to the wall, transfer the supernatant Transfer to another culture dish, change the medium after culturing for 12 hours, and observe the teroid cells under a microscope after continuing culturing for 3-5 days.

[0052] 2. Flow cytometry labeling specific ma...

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Abstract

The invention relates to a human lung teterocyte line construction method, and belongs to the technical field of biological medicine. The method comprises the following steps: transfecting a lentivirus carrying a TERT (telomerase reverse transcriptase) gene and a CDK4 gene into primary human lung telocells; and performing continuous subculture on the human lung teterocyte which is identified to be stably transfected with the TERT and the CDK4 for more than 50 generations to obtain the stable human lung teterocyte line. The teterocyte obtained by the invention can be used for fundamental research on pathogenesis and intervention of acute and chronic diseases, tumors and the like, and can be used for research on intercellular communication and interaction with the teterocyte or other cell types and the like; a cell model is provided for clinical new drug screening and drug treatment mechanism research, and a reserve is provided for subsequent research.

Description

technical field [0001] The invention relates to a method for constructing a human lung-specific cell line, belonging to the technical field of biomedicine. Background technique [0002] The current research on the function of the special cells is to use the special cells extracted from different organs (lung, kidney, heart, etc.) of different species of animals (such as mice, rats, pigs, etc.), the extraction process is time-consuming and expensive every time Extraction requires separation, purification and identification, which requires a lot of manpower and material resources. Since multiple separations and extractions of primary cells are performed by different personnel, it is difficult to guarantee the purity and stability of the extracted cells. How to ensure the purity of cells, and make cells can be passed on for a long time, ensure the stability of cells (avoid batch differences) for clinical application research and basic research, as well as research in the field...

Claims

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Application Information

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IPC IPC(8): C12N15/867C12N5/10C12N5/071C12N15/54
CPCC12N5/0688C12N15/86C12N9/1276C12N9/12C12Y207/11022C12Y207/07049C12N2740/15043C12N2510/04C12N2509/00C12N2800/107
Inventor 宋东莉王向东
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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