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Culture medium and three-dimensional culture method for lung tumor cells

A technology for tumor cells and culture medium, which is applied in the directions of tumor/cancer cells, cell culture active agents, biochemical equipment and methods, etc. It can solve the problems of differences in culture conditions and unpredictability, and achieve the effect of wide use.

Pending Publication Date: 2020-04-24
重庆宇珩生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But the biggest challenge in building organoids of any type in vitro is determining the nutrients, cytokines, and culture methods needed to form them in a dish
These exact culture conditions vary significantly and are unpredictable for different tumor and tissue types

Method used

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  • Culture medium and three-dimensional culture method for lung tumor cells
  • Culture medium and three-dimensional culture method for lung tumor cells
  • Culture medium and three-dimensional culture method for lung tumor cells

Examples

Experimental program
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Effect test

Embodiment 1

[0075] Example 1. Cultivation of Circulating Tumor Cells from Lung Adenocarcinoma Patients Using Organoid Medium

[0076] According to the formula shown in Table 1, an organoid medium for culturing circulating tumor cells from lung cancer patients was prepared according to conventional methods in the art. What is adopted in this embodiment is the specific concentration in column 3 in Table 1, and the source of each component is referred to column 2 in Table 1. Those skilled in the art can understand that although the specific manufacturers and batch numbers of each component are listed in this embodiment, the present invention is not limited to these specific commercially available components, but also includes their equivalents. Column 4 in Table 1 gives the concentration range of each component in the present invention for reference.

[0077] Table 1

[0078]

[0079]

[0080] In the process of adjusting the formula of the experimental medium, the inventors found tha...

Embodiment 2

[0098] Example 2. Using organoid medium to cultivate circulating tumor cells from lung squamous cell carcinoma patients

[0099] Using the same medium and method as in Example 1, a blood sample from a patient with squamous cell carcinoma of the lung was processed, and by enriching and culturing circulating tumor cells in the blood sample, tumor organoids were successfully obtained ( image 3 ).

Embodiment 3

[0100] Example 3. Stemness of cultured cells

[0101] In order to determine whether the cell mass obtained by the three-dimensional culture of the present invention retains the stemness of tumor stem cells, the inventors measured the expression levels of stemness-related genes CD133, CD44 and ABCG2, and compared with those obtained in ordinary two-dimensional (2D) culture The expression levels of related genes were compared in simulated samples cultured in the same medium under the same conditions. The simulated samples were obtained by adding 10 tumor cell line cells (human non-small cell lung cancer cells H2347 cells) to 4ml of peripheral blood. The results are as follows: Figure 5 shown. from Figure 5 It can be seen from the figure that in the cell mass obtained by the three-dimensional (3D) culture method of the present invention, the expression levels of CD133 and ABCG2 are significantly higher than those of ordinary 2D cultured cells, wherein the CD133 expression le...

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Abstract

The invention provides a culture medium for lung tumor cells. The culture medium is particularly suitable for in-vitro three-dimensional culture of lung tumor cells, particularly circulating tumor cells from blood of lung cancer patients. The invention also provides a method for in-vitro three-dimensional culture of the circulating tumor cells to form organoids.

Description

technical field [0001] The invention belongs to the field of cell culture, and relates to a method for culturing circulating tumor cells in vitro and making them differentiate into tumor organoid tissue, and a culture medium used in the method. In particular, the present invention relates to the in vitro culture of circulating tumor cells from lung tumors. Background technique [0002] Circulating tumor cells (circulating tumor cells, CTCs) are formed by some highly invasive and highly metastatic cells in the primary tumor, breaking through the basement membrane and entering the circulatory system, and are closely related to tumor metastasis and recurrence. In recent years, it has been widely reported in various malignant tumors that CTCs are significantly correlated with patient survival and treatment response, especially circulating tumor stem cells (Circulating Tumor Stem Cells, CTSCs) in CTCs, which are important targets for cancer therapy , so CTCs and CTSCs have broad...

Claims

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Application Information

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IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2500/32C12N2500/38C12N2501/02C12N2501/11C12N2501/115C12N2501/119C12N2501/405C12N2501/727C12N2501/998C12N2513/00
Inventor 张文赵长云周敏郑敏周文涵杜菲菲
Owner 重庆宇珩生物科技有限公司
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