Method for separation and in-vitro culture of pancreatic cancer tissue organs of human

An in vitro culture and pancreatic cancer technology, applied in the field of tumor biology, can solve the problems of low transplantation success rate, difficulty in constructing priced PDTXs, etc., and achieve the effect of improving the yield

Active Publication Date: 2019-10-11
SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, the extremely low transplantation success rate and high price make it difficult to construct PDTXs

Method used

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  • Method for separation and in-vitro culture of pancreatic cancer tissue organs of human
  • Method for separation and in-vitro culture of pancreatic cancer tissue organs of human
  • Method for separation and in-vitro culture of pancreatic cancer tissue organs of human

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Embodiment 1

[0072] A method for separating and culturing human tumor tissue organoids in vitro, all separation and culturing operations are performed in a biological safety cabinet, including the following steps:

[0073] (a) Preheat the 24-well plate in a 37°C incubator 6 hours before tissue isolation;

[0074] (b) Melt Human Digestion Medium (digestion solution without DNase I and Y-27632) in a 37°C water bath and bring the temperature to 25-37°C. Each 1g of tissue requires 9ml of Human Digestion Medium for digestion;

[0075] (c) Add 10 μl of 10 mg / ml DNase I and 10 μl of 10.5 mM Y-27632 to 10 ml of Human DigestionMedium (final concentrations of 10 μg / ml and 10.5 μM, respectively);

[0076] (d) Add 3ml of pre-warmed Human Digestion Medium to a 15ml centrifuge tube;

[0077] (e) Record tissue information: human pancreatic cancer tissue (from a hospital in Guangzhou);

[0078] (f) using sterile forceps to transfer the tissue into a petri dish;

[0079] (g) Record the tissue size, shap...

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Abstract

The invention discloses a method for separation and in-vitro culture of pancreatic cancer tissue organs of human. The method comprises the following steps that (1) after the pancreatic cancer tissue of human is cut, enzymic digestion is repeatedly carried out for 10-20 minutes at 37 DEG C in a rotation speed of 30-40 rpm three times, and a pancreatic cancer single cell is obtained; (2) the pancreatic cancer single cell and matrigel are uniformly mixed, then a culture medium containing a growth factor PEG2 and gastrin I are added for culture, and the pancreatic cancer tissue organs of human areobtained. The method for separation and in-vitro culture of the pancreatic cancer tissue organs of human has the advantages that the tumor tissue is digested by adopting a short-time repeated digestion method, which can effectively improve the cell yield and cell viability; the culture medium containing the growth factor PEG2 and the gastrin I is adopted for culture, which can effectively promotethe growth of the pancreatic cancer organs and increase the survival rate of the pancreatic cancer organs.

Description

technical field [0001] The invention relates to the field of tumor biology, in particular to a method for isolating and culturing human pancreatic cancer tissue organoids in vitro. Background technique [0002] Over the past few decades our understanding of primary tumors has expanded and deepened. Although the treatment of some tumors has undergone substantial development, tumor treatment is still a worldwide health problem. The development of new, targeted treatments offers an opportunity to address this problem. Among them, how to transform basic research results into clinical treatment programs is the main bottleneck in the development of new treatment programs. The main reason is that the tumor models used in basic research are difficult to reflect the real situation of patients' tumors. Therefore, although many drugs have good therapeutic effects in tumor models in basic research, they have failed in clinical trials. Although animal tumor models can provide importa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2500/84C12N2501/10C12N2501/345C12N2509/00C12N2509/10C12N2513/00C12N2533/90
Inventor 黄炳培孟亚明陈逸钿孔祥展孟琼赵新保苏良平黄城
Owner SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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