Bionic micro-fluidic chip for simulating in vivo tumor cells and metastasis microenvironment

A microfluidic chip and tumor cell technology, applied in the field of bionic microfluidic chips, can solve the problems of long experimental period, expensive animal models, unable to correctly reflect the physiological state of lung cancer cell metastasis, etc., to ensure gas exchange and easy operation. , the effect of good biocompatibility

Active Publication Date: 2015-08-12
THE SECOND HOSPITAL OF DALIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, currently available animal models for evaluating normal physiological and disease processes are expensive, subject to lengthy experimental cycles, and present various ethical controversies
More importantly, the above-mentioned animal models cannot well control the location and tropism of metastatic lung cancer cells, resulting in the inability to correctly reflect the physiological state of lung cancer cell metastasis in humans.

Method used

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  • Bionic micro-fluidic chip for simulating in vivo tumor cells and metastasis microenvironment
  • Bionic micro-fluidic chip for simulating in vivo tumor cells and metastasis microenvironment
  • Bionic micro-fluidic chip for simulating in vivo tumor cells and metastasis microenvironment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 A bionic microfluidic chip for simulating the microenvironment of lung cancer metastasis in vivo

[0059] The microfluidic chip of the present invention is an airtight whole formed by interlacing and irreversible sealing of three layers of PDMS substrates and two layers of porous PDMS membranes; the first layer of PDMS substrate 1 is provided with channels 11 for air circulation, The inlet 12 and outlet 13 for liquid to enter and exit are respectively located in the upper half of the vacuum channels 14 and 15 on both sides of the channel 11; the second layer of PDMS substrate 2 is provided with a channel 21 for liquid circulation and an inlet 22 for liquid to enter and exit and the outlet 23, respectively located in the lower half of the vacuum channels 14, 15 on both sides of the channel 21, the side of the channel 21 is provided with three connecting channels 211, 212, 213 extending outwards, the connecting channels 211, 212, 213 The ends are respectively pr...

Embodiment 3

[0073] Example 3 Construction of a bionic model simulating the microenvironment of lung cancer cell metastasis in vivo

[0074] 1. 2D culture of lung cancer cells in a microfluidic chip

[0075] (1) The microfluidic chip is sterilized by ultraviolet irradiation, and the porous PDMS membrane is coated with BME. The specific coating process is: pretreatment of the chip, so that cells can better adhere to the porous membrane surface of the chip. Dilute BME (Cultrex basement membrane extract, R&D Systems, McKinley Place, MN, USA) at a ratio of 1:10, mix well, inject into the sample inlet of the microfluidic chip with a micro-sampler, and wait for the gel to solidify overnight in the incubator.

[0076] (2) The microfluidic chip is turned over, that is, the third PDMS substrate faces upward. Collect the suspended mononuclear cells into a centrifuge tube, centrifuge at 1000 rpm for 5 minutes, discard the supernatant, and add fresh medium to prepare a cell suspension. The mononucle...

Embodiment 4

[0086] Example 4 Detection of the effectiveness of the bionic model for simulating the metastatic microenvironment of lung cancer cells in vivo

[0087] Evaluate the effectiveness of the bionic model of the simulation body lung cancer cell transfer microenvironment constructed in Example 3, and complete it through the following experiments:

[0088] 1. Detection of cell viability

[0089] Detection method: absorb the culture medium in the channel in the chip system, inject PBS into the chip channel, and wash the cells of different treatment groups twice; then pump H33342 (1:100) into the staining for 15 minutes, and wash twice with PBS solution; Pump in PI staining (1:200) for 5 minutes, wash with PBS solution twice; under a microscope, observe the fluorescence intensity under the excitation of the corresponding excitation light and record it by taking pictures.

[0090] 2. Detection of co-localization of lung cancer cells and bronchial epithelial cells

[0091] The cell tra...

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Abstract

The invention discloses a bionic micro-fluidic chip for simulating in vivo tumor cells and their metastasis microenvironment. The micro-fluidic chip is formed by interlacing and irreversible sealing of three layers of PDMS substrates and two layers of porous PDMS membranes. The first PDMS substrate layer is provided with a channel for air circulation, an inlet and an outlet for entry and exit of a liquid and an upper half part of a vacuum channel. The third PDMS substrate layer is provided with a cell culture room for cell culture. The channel of the first PDMS substrate layer is positioned up the channel of the second PDMS substrate layer. The cell culture room on the third PDMS substrate layer is positioned down the channel of the second PDMS substrate layer. The micro-fluidic chip provided by the invention can be used for in vitro detection of the tumor cell metastasis process and provides a basis for making clinical tumor treatment plans.

Description

technical field [0001] The invention belongs to the field of clinical application, and relates to a bionic microfluidic chip for simulating the microenvironment of tumors and their metastases in the body. The microfluidic chip of the present invention can dynamically monitor the metastasis process of tumor cells, define the migration mode of tumor cells, and provide guidance for preventing and treating tumor metastasis. Background technique [0002] Cancer cell metastasis is the main cause of death from all cancers, including lung cancer, and is one of the biggest challenges for basic researchers and clinical scholars. Cancer cell metastasis is a complex physiological process that usually includes primary site tumor lesion proliferation, tumor cell detachment, tumor cell migration, extravasation, and formation of metastatic tumor cell clusters at second organs. Tumor cells with a metastatic phenotype exhibit the following properties: enhanced cell mobility, increased abilit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12Q1/02G01N33/68
CPCB01L3/5027B01L3/502707C12M23/16C12M35/08G01N33/5005
Inventor 王琪高占成许志赟郭哲郝华龙徐一彤
Owner THE SECOND HOSPITAL OF DALIAN MEDICAL UNIV
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