Array-type cell dynamic culturing and regionalization processing micro-fluidic chip, and preparation method and application of same

A microfluidic chip, dynamic culture technology, applied in biochemical equipment and methods, tissue cell/virus culture devices, support/immobilization methods for microorganisms, etc. Shear force dynamic loading and other issues to achieve good biocompatibility

Active Publication Date: 2018-04-06
NORTHWESTERN POLYTECHNICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the currently commonly used microfluidic chip realizes the regular renewal of the medium and the timely removal of metabolic waste in the controlled cell culture, all of which are population cell culture and research in a homogeneous environment (Kim S.H., Ahn K., Park J.Y., Responses of human adipose-derived stem cells to interstitial level of extremely low shear flows regarding differentiation, morphology, and proliferation.Lab Chip, 2017, DOI: 10.1039 / c7lc00371d), less able to achieve cell array culture and fluid shear force Dynamic loading and targeted biochemical factor / drug treatment of cells in one or several areas make it more difficult to observe the interaction between treated cells and untreated cells in real time in the same space (Pang L., Liu W ., Tian C., et al., Construction of single-cell arrays and assay of cell drug resistance in an integratedmicrofluidic platform.Lab Chip, 2016, 16, 4612-4620)

Method used

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  • Array-type cell dynamic culturing and regionalization processing micro-fluidic chip, and preparation method and application of same
  • Array-type cell dynamic culturing and regionalization processing micro-fluidic chip, and preparation method and application of same
  • Array-type cell dynamic culturing and regionalization processing micro-fluidic chip, and preparation method and application of same

Examples

Experimental program
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Effect test

Embodiment 1

[0038] An array type cell dynamic culture and regionalized processing microfluidic chip, the chip includes a flow layer 1, a control layer 2, a film layer 3 and a glass layer 4 sealed sequentially from top to bottom; the flow layer 1 consists of an array Cell culture area 5, buffer structure area 6, cell inoculation micropipe 7, left liquid inlet micropipe 8, central liquid inlet micropipe 9 and right liquid inlet micropipe 10, the cell inoculation micropipe 7 is used to flow Layer input cells, the central liquid inlet microchannel 9, the left liquid inlet microchannel 8 and the right liquid inlet microchannel 10 are respectively used as the medium inlet microchannel in the center and the biochemical factor / medicine inlet microchannel on both sides. It is used to provide culture medium for cells cultured in an array to realize fluid shear stress loading and regionalized treatment of biochemical factors / drugs; the arrayed cell culture area 5 is provided with several arrayed arra...

Embodiment 2

[0045] The control layer microvalve 13 drives the liquid in the medium inlet micropipe 9 as follows: Figure 5 , Figure 6 shown. When the micropump 13 was in a quiescent state, the liquid in the medium inlet micropipe 9 was also in a quiescent state, as Figure 5 As shown; when the micropump 13 is driven with a pressure of 2 psi and a frequency of 1 Hz, the three microcavities 18, 19, and 20 of the micropump 13 are sequentially deformed to drive the liquid flow in the medium inlet micropipe 9, as Figure 6 As shown, so as to achieve real-time update of the medium.

Embodiment 3

[0047] This embodiment takes the array culture of osteoblast-like cells IDG-SW3 in a microfluidic chip as an example.

[0048] Step 1. Pretreating the microfluidic chip. First, the prepared microfluidic chip in Example 1 was sterilized by ultraviolet irradiation, and then the fluid layer microchannel and cell culture area were protein-coated with type I collagen, 0.15mg / mL, 37°C, 2h, Then use serum-free α-MEM medium to rinse the fluidized layer microchannel and cell culture area.

[0049] Step 2. Prepare osteoblast-like cell IDG-SW3 suspension. Culture IDG-SW3 cells by conventional cell culture method, digest with trypsin to obtain cells in the logarithmic growth phase, centrifuge the cell suspension at 800 rpm for 5 minutes to remove trypsin, resuspend the cells in α-MEM medium containing 10% fetal bovine serum, The cell density was adjusted to 0.5×10 6 cells / mL.

[0050] Step 3, inoculating cells. Drive the micropump 15 just below the main pipeline 11 with 2psi gas pres...

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Abstract

The invention discloses an array-type cell dynamic culturing and regionalization processing micro-fluidic chip which includes, in a manner of being successively encapsulated from top to bottom, a flowing layer, a control layer, a thin film layer and a glass layer. The flowing layer is composed of an array-type cell culture zone, a buffer structure zone, a cell inoculation micro-pipe, a left liquidfeeding micro-pipe, a central liquid feeding micro-pipe and a right liquid feeding micro-pipe; the array-type cell culture zone is provided with a plurality of array-type U-shaped culture grooves anda plurality of U-shaped micro-columns are disposed in the buffer structure zone; the control layer is composed of a plurality of micro-pumps in a manner that an end-sealed structure is composed of one inlet and a plurality of micro-cavities in series connection. The micro-fluidic chip is used for carrying out fluid shear stress dynamic loading and regionalization processing of more than two biochemical factors/medicines, and researching interaction between processed cells and non-processed cells. The micro-fluidic chip breaks through conventional cell culture and treatment modes, and has beneficial to simulation of a fluid micro-environment and a biochemical micro-environment in cells in tissue under different physiological or pathological conditions.

Description

technical field [0001] The invention relates to the technical field of an array cell culture microfluidic chip, in particular to an arrayed cell dynamic culture and regionalized treatment microfluidic chip and its preparation method and application. Background technique [0002] Cell culture based on microfluidic chips is an important technical means of life science research, which is conducive to obtaining rich information on cell structure and function in a simple and controllable environment compared with complex systems in vivo. However, the currently commonly used microfluidic chip realizes the regular renewal of the medium and the timely removal of metabolic waste in the controlled cell culture, all of which are population cell culture and research in a homogeneous environment (Kim S.H., Ahn K., Park J.Y., Responses of human adipose-derived stem cells to interstitial level of extremely low shear flows regarding differentiation, morphology, and proliferation.Lab Chip, 2...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12Q1/02
CPCC12M23/16C12M25/04G01N33/5008
Inventor 任丽商澎王圣航武婉情叶芳杨鹏飞王哲龚翰林
Owner NORTHWESTERN POLYTECHNICAL UNIV
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