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Serum-free medium for liver cancer stem cells and culture method for serum-free medium

A serum-free medium, liver cancer stem cell technology, applied in the direction of tumor/cancer cells, animal cells, vertebrate cells, etc., can solve the problems of difficult separation, small number of stem cells, difficult to carry out, etc., and achieves high spheroidization rate and abundant experiments. effect of material

Inactive Publication Date: 2015-12-09
广西壮族自治区肿瘤医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] More and more people agree with the existence of tumor stem cells, but the number of stem cells is too small to be separated, so it is difficult to carry out follow-up experiments

Method used

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  • Serum-free medium for liver cancer stem cells and culture method for serum-free medium
  • Serum-free medium for liver cancer stem cells and culture method for serum-free medium
  • Serum-free medium for liver cancer stem cells and culture method for serum-free medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1: Hepatoma cell line MHCC-97H stem cell culture

[0054] 1. Culture medium preparation: Add 2ug each of EGF, FGF-10, IGF-1, 2ml of B27, 5g of heparin, 1ml of penicillin + streptomycin double antibody solution to 96.9ml of DMEM / In F12+GlutaMAXTM-1 medium, pipette to mix.

[0055] 2. Resuscitation of the human liver cancer cell line MHCC97H: Take the cells out of the -80°C liquid nitrogen tank, immediately shake them gently in a 37°C water bath for about 2 minutes, and make them fully and evenly heated and then melt quickly. In the ultra-clean workbench, add 5ml of high-sugar DMEM complete medium containing 10% FBS (containing penicillin + streptomycin double-antibody solution) and gently blow and mix to neutralize DMSO, centrifuge at 1500rpm for 5min, and discard the supernatant . Resuspend the cells in high-sugar DMEM complete medium (containing penicillin + streptomycin double-antibody solution) containing 10% FBS, pipette and mix well, and use a disposable...

Embodiment 2

[0063] Example 2 Hepatoma cell line MHCC-97L stem cell culture

[0064] MHCC-97L is donated by Fudan University Medical Center

[0065] 1. Reagent preparation: Add 2ug each of EGF, FGF-10, IGF-1, 2ml of B27, 5g of heparin, 1ml of penicillin + streptomycin double antibody solution to 96.9ml of DMEM / F12+GlutaMAXTM in a clean bench -1 Medium, mix by pipetting.

[0066] 2. Resuscitate the human liver cancer cell line MHCC97L: Take the cells out of the -80°C liquid nitrogen tank, immediately shake them gently in a 37°C water bath for about 2 minutes, make them fully and evenly heated and then melt quickly. In the ultra-clean workbench, add 5ml of 10% FBS high-sugar DMEM complete medium (containing penicillin + streptomycin double antibody solution) and gently blow and mix to neutralize DMSO, centrifuge at 1500rpm for 5min, and discard the supernatant. Then resuspend the cells with 10% FBS high-sugar DMEM complete medium (containing penicillin + streptomycin double antibody soluti...

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Abstract

The invention provides a serum-free medium for liver cancer stem cells and a culture method for the serum-free medium. Based on a DMEM / F12+GlutaMAXTM<-1> medium, the serum-free medium comprises a recombinant human epidermal growth factor, a recombinant human basic fibroblast growth factor-10, an insulin No.1 growth factor, a serum-free complementary nutritional ingredient, heparin and a penicillin and streptomycin double-resistant solution. The serum-free medium has the effects as follows: 1, three special growth factors are added to the serum-free medium in a corresponding proportion; direct serum-free culture is realized, so that liver cancer cells can be pelletized, the pelletizing rate is high, and continuous and stable passage can be realized; 2, the growth factors in the serum-free medium keep a small group of cells in an undifferentiated state; the serum-free medium and the culture method have wide prospect in fundamental research and clinical application of serum-free culture of the liver cancer stem cells and provide rich experiment materials for research of the liver cancer stem cells.

Description

technical field [0001] The invention belongs to the technical field of biological cell culture, and in particular relates to a serum-free culture medium of liver cancer stem cells and a culture method thereof. Background technique [0002] In recent years, theoretical research on cancer stem cells has received widespread attention and is considered to be the most mainstream reason. Michor et al. believe that stem cells are a small group of cells that have self-renewal, unlimited proliferation, and multidirectional differentiation in the original cells (HwangHI, LeeTH, JangYJ. Cell proliferation-inducing protein52 / Mitofilinisa surfaceantigenonundifferentiatedhumandentalpulpstemcells[J].StemCellsDev, 2015). However, cancer stem cells (cancer stem cells, CSCs) also exist in tumor cells, and have strong self-renewal and unlimited proliferation capabilities in terms of tumorigenic potential, and also play a role in the occurrence, development, metastasis, invasion, and recurrence...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/095
Inventor 黎乐群马良游雪梅李航向邦德钟鉴宏彭榆翀陆世东欧超陈洁齐鲁楠龚文锋
Owner 广西壮族自治区肿瘤医院
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