Isolation and culture method of primary cervical cancer cells

Abstract

The invention discloses an isolation and culture method of primary cervical cancer cells. The isolation and culture method of the primary cervical cancer cells comprises the following steps: 1) treating a tissue block in a pre-cooled sterile culture medium; 2) cleaning tissues, and cutting-up the cleaned tissues; 3) carrying out digestion treatment; 4) carrying out filtration by using an aseptic cell filter so as to obtain a filtered digestion solution, and carrying out low-temperature storing; 5) rinsing the aseptic cell filter with collagenase, and carrying out collection by using a centrifuge tube; 6) placing the centrifuge tube flat in an oscillation box so as to be subjected to full oscillation to obtain a centrifuged digestion solution; 7) combining the filtered digestion solution obtained in the step 4) and the centrifuged digestion solution obtained in the step 6), carrying out centrifuging, and discarding liquid supernatant; 8) carrying out full blowing so as to allow resuspension of cell sediments; 9) carrying out isolation and passage; and 10) carrying out magnetic-activated cell sorting. The isolation and culture method of the primary cervical cancer cells has the following beneficial effects: the method is capable of prolonging tissue in-vitro time, improving cell growth efficiency, shortening time cycle and ensuring high purification efficiency; and thus, a noveland efficient solution is provided for basic study of cervical cancer.

Description

technical field [0001] The invention relates to the field of modern biotechnology, in particular to a method for separating and culturing primary cells of cervical cancer tumors. Background technique [0002] Worldwide, cervical cancer (CC) is the second most common female malignancy after breast cancer. It is well known that persistent high-risk HPV infection is the cause of the disease. Reduce the prevalence of CIN ≥ III by at least 50% (American Society of Clinical Oncology report, 2016). However, due to differences in mass screening and primary prevention, and lack of effective HPV+ regression strategies, cervical cancer is still the most important fatal disease of female reproductive system tumors. According to an article written by the National Cancer Center of China (Cancer statistics in China, 2015), the incidence of cervical cancer is as high as 99 / 100,000 people, and the death rate is 31 / 100,000 people, which seriously threatens the physical and mental health of C...

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Problems solved by technology

[0003] At present, cervical cancer cell lines include the widely used Hela (derived from cervical adenocarcinoma), and SiHa, CaSki, C4-1 and C33A derived from cervical squamous cell carcinoma. Compared with other cancer cell lines, there are fewer types and cannot fully satisfy required for research

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