Continuous culture method and culture medium for primary hepatocellular carcinoma organoids

A technology for culture medium and hepatocytes, which can be used in cell culture active agents, hepatocytes, detection of programmed cell death, etc., can solve the problems of low culture success rate, low success rate, and difficulty in culture

Pending Publication Date: 2021-12-17
上海诺典生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] There are very few reports on the successful culture of hepatocellular carcinoma organoids. In 2017, Nature medicine (NM) reported 3 cases of hepatocellular carcinoma organoids were successfully cultured and there were no follow-up reports. Domestic scientific research institutions also reflect that primary hepatocellular carcinoma organoids continue to be cultured However, the success rate is extremely low (5%-10%). The fundamental reason is that the intertumoral and intratumoral heterogeneity of hepatocellular carcinoma is greater than that of other tumors, resulting in a single hepatocellular carcinoma organoid medium culture success rate. low, and therefore a diverse medium is critical for the successful culture of hepatocellular carcinoma organoids

Method used

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  • Continuous culture method and culture medium for primary hepatocellular carcinoma organoids
  • Continuous culture method and culture medium for primary hepatocellular carcinoma organoids
  • Continuous culture method and culture medium for primary hepatocellular carcinoma organoids

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1, improved culture medium 1

[0044] DMEM / F12 medium, ascorbic acid (10mg / L), L-glutathione (1-50mg / L), Insulin (20mg / L), Sodium selenite (0.05mg / L), R-spondin 1 (500ng / ml), N2(1X), B27(1X), GlutaMAXSupplement(1X), HEPES(1X), Niacinamide(20mM), Normocin TM (1X), N-Acetylcysteine ​​(2mM), A83-01 (10μM), Gastrin I (10nM), EGF (20ng / ml), HGF (50ng / ml), FGF7 (20ng / ml) , FGF10 (200 ng / ml), Y27632 (50 μM).

Embodiment 2

[0045] Embodiment 2, improved culture medium 2.

[0046] DMEM / F12 medium, ascorbic acid (10mg / L), Holo-Transferrin (50mg / L), L-glutathione (20mg / L), ethanolamine hydrochloride (30mg / L), R-spondin 1:( 200ng / ml), N2(1X), B27(2X), GlutaMAXSupplement(1X), HEPES(1X), Nicotinamide(5mM), Normocin TM (1X), N-Acetylcysteine ​​(2mM), A83-01 (5μM), Gastrin I (10nM), EGF (50ng / ml), FGF7 (100ng / ml), FGF10 (100ng / ml) , DMSO (1%), Y27632 (50 μM).

Embodiment 3

[0047] Embodiment 3, improved culture medium 3

[0048] DMEM / F12 medium, Holo-Transferrin (10mg / L), L-glutathione (1-10mg / L), Insulin (10mg / L), R-spondin 1: (100ng / ml), N2 (0.5 X): B27(1X), GlutaMAX Supplement(1X), HEPES(1X), Nicotinamide(10mM), Normocin TM (1X), N-acetylcysteine ​​(1.25 mM), A83-01 (0.5 μM), gastrin I (1 nM), EGF (100 ng / ml), Y27632 (50 μM), CHIR99021 (50 μM).

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Abstract

The invention discloses a culture medium for primary liver cancer organoids and a culture method of the primary liver cancer organoids. By adjusting the contents of various components in the culture, the organoid culture of the primary hepatocellular carcinoma cells with huge heterogeneity can be realized, and a foundation is laid for revealing the pathogenesis of hepatocellular carcinoma and screening anti-cancer drugs.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a medium for primary liver cancer organoids and a culture method thereof. Background technique [0002] Primary liver cancer (primary liver cancer) is one of the most common malignant tumors in the world, ranking second among all malignant tumor-related causes of death in the world. China is the country with the highest incidence and mortality of liver cancer, and the number of new cases of liver cancer every year Both cancer and death toll account for 50%-55% of the world, and it is the second cancer killer in my country. Primary liver cancer includes three different pathological types: hepatocellular carcinoma, intrahepatic cholangiocarcinoma and mixed type, among which hepatocellular carcinoma accounts for more than 85%-90%. The long-term primary 2D culture of hepatocellular carcinoma in vitro is difficult and the success rate is low, and the successfully established 2D hepatocellu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/09C12Q1/02
CPCC12N5/0671C12N5/0693G01N33/5011C12N2513/00C12N2500/38C12N2500/32C12N2501/11C12N2501/345C12N2500/46C12N2501/117C12N2501/119C12N2500/25C12N2500/62C12N2500/60C12N2503/02G01N2500/10
Inventor 徐小雅
Owner 上海诺典生物科技有限公司
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