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A human bronchial epithelial cell line hbe-tt

A technology of bronchial epithelium and cell lines, applied in the biological field, can solve problems affecting the accuracy of experimental data

Active Publication Date: 2018-08-24
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this characteristic of SV40 virus gene is only limited to the early stage of cell line establishment. With the increase of in vitro passage times, these immortalized cell lines or cell lines will also show certain malignant characteristics, and these cell characteristics will affect Accuracy of experimental data

Method used

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  • A human bronchial epithelial cell line hbe-tt
  • A human bronchial epithelial cell line hbe-tt
  • A human bronchial epithelial cell line hbe-tt

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Preparation of human bronchial cell line HBE-TT

[0028] 1.1 Acquisition and culture of primary bronchial epithelial cells

[0029] Source of bronchial tissue: normal tissue next to diseased tissue collected through thoracotomy before treatment. The patient is male, Han nationality, diagnosed with lung cancer, understands and understands the purpose and content of the research, and signs an informed consent. The sample collection was approved by the Ethics Committee of Sun Yat-sen University, and the discarded samples were handled in strict accordance with the principles and procedures of biological pollutant treatment.

[0030] The obtained human bronchial normal tissues were immediately preserved in pre-cooled DMEM medium and brought back to the laboratory within half an hour. The mucus and congestion in the lumen were washed with cold PBS containing double antibiotics (penicillin / streptomycin dual antibiotics), the connective tissue was removed, and the...

Embodiment 2

[0044] Example 2 Detection of LT protein expression in human bronchial cell line HBE-TT

[0045] The human bronchial cell line HBE-TT, which is a positive cell screened in Example 1, was further identified by Western blotting.

[0046] In this experiment, a cell line with high expression of SV40LT was used as a positive control for detecting LT. The cell line used for the positive control only needs to have the characteristic of high expression of SV40LT. Existing cell lines with this characteristic can be used as the positive control. Human bronchial epithelial cells from the Type Culture Collection ATCC, catalog number NL20 CRL2503 TM , the human bronchial epithelial cells highly expressing the SV40LT gene can also be used as a positive control. Specifically used in this example are HEK cells with high expression of SV40LT gene preserved in our laboratory (Hahn WC, DessainSK, Brooks MW, King JE, Elenbaas B, Sabatini DM, DeCaprio JA, WeinbergRA. Enumeration of the simian v...

Embodiment 3

[0049] Example 3 Cell Growth Characteristics

[0050] The human bronchial cell line HBE-TT cells were inoculated into 24-well plates, and each well was inoculated with 5×10 4 cells, set up 3 parallel samples, and the culture medium used was human bronchial epithelial cell culture medium, and counted at 48h, 72h, and 96h. The cell growth curve was drawn with the observation days as the abscissa and the cell number as the ordinate. Calculate the doubling time. The cell doubling time refers to the time for the number of cells to double, which can better reflect the growth status of the cells. Cell doubling time was calculated using the following formula. where TD is the cell doubling time, N t Indicates the number of cells on day t, N 0 Indicates the number of initially inoculated cells, and Δt indicates the days from inoculation to counting.

[0051]

[0052] Take the culture time as the X-axis and the cell number as the Y-axis to draw the cell growth curve, see Figure...

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Abstract

The invention a novel human bronchial epithelial cell strain HBE-TT, collected in China Center for Type Culture Collection (Wuhan) under CCTCC NO: C201560. The cell strain HBE-TT is verified to be good in purity, free of sundry cell contamination and differed from all existing other cell strains. In addition, the immortalized cells retain typical features of primary bronchial epithelial cells and are free of mycoplasma contamination and vicious transformation features, and it is suggested that cells HBE-TT can retain trait stability during in-vitro subculture. The cells HBE-TT are expected to be a forcible tool for the deep discussion on occurrence and development molecular mechanism of lung diseases caused by environmental toxic factor exposure and for pharmacological study, and the cells are significant to the study on health damage effect of toxic factors on human lung as well as risk estimation and new drug development.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to an immortalized human bronchial epithelial cell line. Background technique [0002] Human bronchial epithelial cells (HBE) are the first barrier between the human body and the external environment, and play an important role in the occurrence and development of various lung diseases caused by environmental harmful factors, such as lung cancer, asthma, Chronic obstructive pulmonary disease, cystic fibrosis, etc. Human bronchial epithelial cells cultured in vitro are similar to the original tissue in vivo in terms of morphology, structure and biological function. Human cell culture technology plays a key role in the development of in vitro research. Since primary cells will senescence and die after being cultured in vitro for a certain period of time, immortalized cell lines must be established. The construction of immortalized human bronchial epithelial cell lines provides very favo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10
Inventor 李道传陈雯邢秀梅
Owner SUN YAT SEN UNIV
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