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Three-dimensional cell chip based on cell printing and multi-parameter sensing array integrated technology

A cell chip, cell technology, applied in the field of biological information, can solve the problems of limiting real-time detection, cell damage, cell death, etc.

Active Publication Date: 2012-03-21
REGENOVO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The classic neural electrophysiological detection method uses microelectrode signal acquisition or confocal fluorescence imaging to record intracellularly. Causes certain damage to cells and leads to cell death in a short period of time, which limits the real-time detection of action potential and ion channel current recording; at the same time, the number of cells that can be selected to collect signals is small

Method used

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  • Three-dimensional cell chip based on cell printing and multi-parameter sensing array integrated technology
  • Three-dimensional cell chip based on cell printing and multi-parameter sensing array integrated technology
  • Three-dimensional cell chip based on cell printing and multi-parameter sensing array integrated technology

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1 cardiomyocyte culture

[0057] Cardiomyocytes derived from neonatal rats were selected as one of the seed cells forming the basis of the system.

[0058] (1) Take one suckling mouse at a time, put it on a piece of sterile gauze, scrub its chest and abdomen with 70% ethanol, remove the heart and put it into the beaker marked 1. Each dissection should not exceed 1 minute.

[0059] (2) Use a hemostat to remove the connective tissue and blood clots on the heart, then transfer it to the beaker marked 2, wash it again, and transfer it to the beaker marked 3. Use a straw to gently suck out the D-Hanks solution in beaker 3, then add 2ml of 0.25% trypsin solution (37°C), use ophthalmic scissors to cut the heart into sand-sized pieces, add 2ml of trypsin solution, and mix well After digestion at 37°C for 15 minutes.

[0060] (3) After digestion, suck off the supernatant, add 4ml of new trypsin solution, mix well, digest at 37°C for 15 minutes, then discard the sup...

Embodiment 2

[0066] Embodiment 2 adrenal chromaffin cell culture

[0067] (1) PC-12 mouse adrenal pheochromocytoma cell line (ATCC CRL-1721, American Type Culture Collection, American Type Culture Collection Center), with high sugar DMEM (Dulbecco's Modified Eagle's Medium, Sigma) + 10% fetal bovine Serum culture medium, cultured at 37°C carbon dioxide constant temperature culture, generally about three days, the cells reach monolayer confluence, after the cells are confluent, carry out routine subculture;

[0068] (2) Subculture: Absorb the culture medium, add 1 mL of 0.2% trypsin solution, quickly wash the bottle wall and then absorb it, then add 1 mL of 0.2% trypsin solution, incubate at 37°C for about 5 minutes, observe under a microscope, and wait for 80% of the After the cells become round, gently absorb the trypsin solution, and quickly add DMEM medium containing 15% FCS to stop the digestion; blow the wall of the culture flask to make a cell suspension again; adjust the cell concen...

Embodiment 3

[0070] Embodiment 3 Design and manufacture of multi-sensor integrated chip

[0071] In the processing technology, both the MEA and ECIS units use gold electrodes and the base structure and protective layer structure are the same, so the processing technology can be shared. For specific processing procedures, see image 3 shown in each step. The processing process takes the silicon substrate as an example. The processing process is briefly described as follows:

[0072]First, the insulating layer on the surface of the device is grown and deposited, and a layer of 100nm Si3N4 is deposited on it by chemical vapor deposition (PECVD) process. Then process multiple functional units of MEA and ECIS. On the basis of the previous steps of processing, a layer of SiO2 with a thickness of 600nm is deposited on the chip by PECVD. The metal layer adopts Au / Cr composite layer, and Cr is used to enhance the adhesion between gold and the silicon base layer on which SiO2 is deposited. , and...

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PUM

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Abstract

The invention relates to a cell chip technology based on cell printing technology and multi-sensor integrated chip technology and application of the cell chip technology in high content medicine screening. An integrated cell MEA (Microelectrode Array) is constructed on glass or a silicon substrate by utilizing micro-electronics processing technology, and an ECIS (Electric Cell-Substrate Impedance System) comprises a disc type ECIS and an IDA (Interdigitated Array). By an arrayed chip of three sensing units, different cells and special imitated extracellular substrate material are mixed, and are positioned and printed at the assigned position in the MEA by a multi-nozzle cell printing device so as to construct the cell chip. A cell sensor on the chip is used for transmitting and recording the static and dynamic parameters such as cell action potential frequency, amplitude, wave form, network signal transmitting speed, cell attaching, moving state and the like. The invention provides a method applying the three-dimensional cell chip for medicine screening.

Description

technical field [0001] The invention belongs to the technical field of biological information, and specifically relates to a new multi-cell printing technology and multi-parameter sensing array integrated chip technology to construct a three-dimensional cell chip, a construction method and its application in high-content drug screening. Background of the invention [0002] The combination of life science, information science and manufacturing science is an important trend in the development of science and technology in the 21st century. Cell 3D printing technology based on advanced manufacturing theory and tissue engineering theory, and cell chip technology based on micro-processing technology and sensor technology have expanded new theoretical and technical space for research fields such as tissue engineering, drug screening, and environmental monitoring. [0003] The three-dimensional cell assembly technology based on computer 3D modeling and advanced manufacturing discret...

Claims

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Application Information

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IPC IPC(8): C12M1/00C12M1/34C12Q1/02
Inventor 徐铭恩徐莹胡金夫葛亚坤郭淼
Owner REGENOVO BIOTECH
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