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Neurofibroma cell line and construction method of cell line

A neurofibromatosis, construction method technology, applied in the direction of tumor/cancer cells, animal cells, vertebrate cells, etc., can solve the problem of difficult long-term survival, lack of neurofibromatosis cell lines, neurofibromatosis cells do not have immortality, etc. problems, to achieve a good effect of reference and promotion

Active Publication Date: 2020-12-29
SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Abstract
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  • Claims
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Problems solved by technology

[0003] However, neurofibroma is a benign tumor. In vitro studies, unlike most malignant tumors, neurofibroma cells do not possess immortality, and it is difficult to survive for a long time in vitro, making it difficult to carry out research
At present, the internationally recognized neurofibroma-derived cell lines are all from abroad, and can be queried and purchased on ATCC (https: / / www.atcc.org). There is a lack of neurofibroma cell lines for Chinese genetic background

Method used

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  • Neurofibroma cell line and construction method of cell line
  • Neurofibroma cell line and construction method of cell line
  • Neurofibroma cell line and construction method of cell line

Examples

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Embodiment 1

[0049] Example 1 Neurofibroma cell line and its construction

[0050] In this example, a neurofibroma cell line is provided, especially a neurofibroma cell line derived from human neurofibromatosis type I, named PNF1-9HOSP-01, and the preservation number of the depository is CCTCC NO :C202006.

[0051] The construction method of the neurofibroma cell line is to transfect the telomerase reverse transcriptase gene (TERT) and CDK4 / m antigen to the neurofibroma cell line isolated in the first generation, and obtain the immortalized neurofibroma cell line through monoclonal screening . And, described construction method comprises the following steps:

[0052] Step S11: Place the sheared neurofibroma tissue into a solution containing 10% (v / v) fetal bovine serum, 1.25U / ml dispase, 300U / ml collagenase, 100U / ml penicillin and 100ug / ml streptomycin After overnight at 37°C, filter through a 30 μm cell strainer to collect the cells, coat the cells with laminin and place them in a me...

Embodiment 3

[0102] Example 3 Verification Results

[0103] In this example, the verification results of the neurofibroma cell line PNF1-9HOSP-01 of the present invention obtained according to the above verification method are provided.

[0104] 2..1 Morphology

[0105] 2.1.1 General shape: such as figure 2 As shown, many newborn cells can be seen, and the cells grow in clusters, with irregular shapes, tight arrangements, clear boundaries, and loss of contact inhibition.

[0106] 2.1.2 Light mirror: such as figure 2 As shown, the cultured living cells were observed under an inverted phase-contrast microscope, most of them were polygonal, with abundant cytoplasm, large and round nuclei, multiple and obvious nucleoli, and mitotic figures were visible.

[0107] 2.1.3 Electron microscope: such as Figure 3A , 3B As shown in and 3C, SEM showed that the cell surface was wrinkled and surrounded by microvilli or protrusions.

[0108] 2.2 Cell dynamics

[0109] 2.2.1 Drawing of cell grow...

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Abstract

The invention provides a neurofibroma cell line, and particularly relates to a neurofibroma cell line derived from human I-type neurofibromatosis and a construction method of the neurofibroma cell line. The neurofibroma cell line has a genetic background of Chinese people and has immortality.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a neurofibroma cell line and its construction method and application. Background technique [0002] Neurofibromatosis type 1 (NF1) is an autosomal dominant genetic disease caused by mutations in the NF1 gene. The prevalence of NF1 in newborns is about 1 / 3500~1 / 3000 (American Journal of Epidemiology, 2000, 151(1):33-40; Journal of Neuroscience History, 2010,19(4):333- 348). Neurofibromas are one of the most common and characteristic symptoms of NF1. Neurofibromas contain a large number of proliferating Schwann cells and other nerve fiber supporting components, such as perineurial cells, fibroblasts, blood vessels, and infiltrating cells. Mast cells. Neurofibromas are mainly divided into cutaneous neurofibromas and plexiform neurofibromas. Skin neurofibromas do not have the potential for malignant transformation, while plexiform neurofibromas mostly show invasive growth, des...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N15/867
CPCC12N5/0693C12N15/86C12N2510/04C12N2740/15043
Inventor 王智超李青峰任捷艺顾熠辉王薇
Owner SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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