Method for co-culturing pluripotent stem cell-derived human liver and pancreas islet based on multiple organ chips

A technology for pluripotent stem cells and cell culture, applied in the field of designing stem cells and tissue engineering, can solve the problems of difficulty in constructing three-dimensional multi-components, limitations, etc., and achieve the effects of being conducive to the maintenance of cell in vitro functions, less consumption, and broad application prospects.

Pending Publication Date: 2021-05-28
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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Problems solved by technology

However, existing research results have shown that the simple mixed culture of liver and islet cells has great limitations on the study of simple islets and liver function, and traditional two-dimensional co-culture such as Transwell is difficult to construct three-dimensional multi-component liver and islet organoids

Method used

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  • Method for co-culturing pluripotent stem cell-derived human liver and pancreas islet based on multiple organ chips
  • Method for co-culturing pluripotent stem cell-derived human liver and pancreas islet based on multiple organ chips
  • Method for co-culturing pluripotent stem cell-derived human liver and pancreas islet based on multiple organ chips

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Embodiment 1

[0047] The microfluidic chip designed and produced by the laboratory is used to construct the co-culture chip of human liver and pancreatic islet organoids derived from pluripotent induced stem cells. The configuration is as follows: figure 1 shown. The microfluidic chip is mainly composed of the microfluidic chip. The chip includes structured PDMS 1 on the upper layer and structured PDMS 2 on the lower layer, connected by cell inlet pools 3, 4, 5, 6, 7, 8, 9, 10, and cell culture chambers. The channel 11 is composed of cell culture chambers 12, 13, 14, 15, 16, 17, 18, 19, and is connected with a peristaltic pump through a perfusion tube 20 and an interface valve 21. In the co-cultivation experiment, the induced islet cell microspheres were resuspended in the medium containing DF12 supplemented with 1% B27, 1% N2, 1% non-essential amino acids, 1% GLUTAMAX, and the final concentration was 10mM glucose. 3 The density per milliliter was inoculated in cell culture chambers 3, 8, ...

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Abstract

The invention relates to a method for co-culturing pluripotent stem cell-derived human liver and pancreas islet based on multiple organ chips. The multi-organ culture chip can be composed of two or more cell culture chambers, the bottom of each culture chamber is provided with a micro-pit (column)-shaped structure arranged in an array mode so that three-dimensional growth of cells can be facilitated, the cell culture chambers are connected in series through a plurality of channels which are not limited in length and micron in width, and a closed continuous perfusion culture system is achieved through a peristaltic pump. Liver and pancreas islet for co-culture are human-derived liver-like and pancreas islet-like at a three-dimensional tissue level induced by pluripotent stem cells and have partial physiological functions of the liver and pancreas islet through gene, protein and secretion level identification, and in the same system, pancreas islet cells and liver cells are mutually promoted through secreted cell factors, co-culture of the pancreas islet and the liver for a long time in vitro and glucose homeostasis regulation closer to a physiological level are realized.

Description

technical field [0001] The present invention designs the field of stem cells and tissue engineering, and is a multi-organ chip for co-culture of human liver and islets derived from pluripotent stem cells, and specifically relates to blood sugar construction of a co-culture system of liver and islet organoids derived from stem cells based on organ chip technology method. Background technique [0002] In recent years, the incidence of diabetes has risen sharply, and it has become the third largest chronic disease in the world, seriously threatening social health. Diabetes can be divided into type 1 diabetes caused by insulin deficiency and type 2 diabetes caused by insulin resistance according to its pathogenesis. The rise in blood sugar caused by the imbalance of sugar regulation in the body causes a series of diabetic syndromes that seriously affect human health. At present, pancreas or islet transplantation is the main method for the fundamental treatment of diabetes, but...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12N5/071
CPCC12M21/08C12M23/16C12M23/34C12N5/0671C12N5/0677
Inventor 秦建华陶婷婷陈雯雯
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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