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CN-122012236-A - Microfluidic-mediated static and dynamic collaborative cell culture chip and application thereof

CN122012236ACN 122012236 ACN122012236 ACN 122012236ACN-122012236-A

Abstract

The invention relates to a microfluidic mediated static and dynamic collaborative cell culture chip and application thereof, relates to the technical field of biotechnology, and aims to solve one of the problems that static culture is difficult to be repeated in-vivo real mechanical microenvironment, a chip device capable of synchronously implementing static and mechanical stretching dynamic biphase culture is lacking in the prior art, and the like. The invention relates to a microfluidic mediated static and dynamic collaborative cell culture chip and application thereof, comprising a cell culture chip and a pneumatic system matched with the cell culture chip, wherein the cell culture chip comprises a microfluidic regulating cavity and a culture cavity, and the pneumatic system regulates the air pressure in the microfluidic regulating cavity to drive the local movement of the bottom surface of the culture cavity so as to enable cells to be cultured to be dynamically cultured and statically cultured in the culture cavity simultaneously. The invention can synchronously develop static and dynamic culture in the same chip, effectively re-etch the internal mechanical microenvironment and synchronously implement static and mechanical stretching dynamic biphase culture.

Inventors

  • JING RAN
  • CUI MING

Assignees

  • 北京大学第三医院（北京大学第三临床医学院）

Dates

Publication Date
20260512
Application Date
20260212
Priority Date
20260119

Claims (10)

  1. 1. The micro-fluidic mediated static and dynamic collaborative cell culture chip is characterized by comprising a cell culture chip (1) and a pneumatic system (2) matched with the cell culture chip (1), wherein the cell culture chip (1) comprises a culture layer (11), a transmission layer (12) and a micro-fluidic bottom layer (13), the culture layer (11) is provided with a culture chamber (111), the culture chamber (111) comprises a static culture area (1111) and a dynamic culture area, and the micro-fluidic bottom layer (13) is provided with a micro-fluidic regulation chamber (131) communicated to the pneumatic system (2); The bottom surface of the culture layer (11), the transmission layer (12) and the top layer of the microfluidic bottom layer (13) are attached to form a whole in a plasma bonding mode, the pneumatic system (2) adjusts the air pressure in the microfluidic regulating and controlling cavity (131), and the transmission layer (12) drives the local movement of the bottom surface of the culture cavity (111) to enable cells to be cultured to be dynamically cultured and statically cultured in the culture chamber at the same time.
  2. 2. The microfluidic mediated static and dynamic co-cell culture chip according to claim 1, wherein the culture chamber (111) is arranged opposite the microfluidic control chamber (131).
  3. 3. The microfluidic mediated static and dynamic co-cell culture chip according to claim 2, wherein the width of the culture chamber (111) is larger than the width of the microfluidic control chamber (131), and the length of the culture chamber (111) is smaller than the length of the microfluidic control chamber (131).
  4. 4. The microfluidic mediated static and dynamic co-cell culture chip according to claim 1, wherein the transmission layer (12) is made of super-elastic PDMS membrane material.
  5. 5. The microfluidic mediated static and dynamic co-cell culture chip according to claim 1, wherein the bottom surface of the culture layer (11) and the top layer of the microfluidic bottom layer (13) are both made of PDMS.
  6. 6. A microfluidic mediated cell static and dynamic co-cultivation method, which is characterized in that the cell culture chip (1) according to any one of claims 1-5 is used for carrying out static and dynamic co-cultivation on cells.
  7. 7. The method according to claim 6, comprising the steps of: step one, paving cells to be cultured on a culture chamber (111); Step two, changing the air pressure value in the micro-flow regulating cavity (131) by utilizing the pneumatic system (2) to increase or decrease the air pressure in the micro-flow regulating cavity (131); And thirdly, the top layer of the micro-flow regulating cavity (131) drives a dynamic culture area (1112) of the culture cavity (111) to move upwards or downwards through the transmission layer (12), so that cells laid in the dynamic culture area (1112) are subjected to maintenance traction stimulation culture, and meanwhile, cells laid in a static culture area (1111) are subjected to static culture.
  8. 8. The method according to claim 7, wherein in the second step, the pneumatic system (2) reduces the air pressure in the micro-fluidic control chamber (131) by 2kPa relative to the atmospheric pressure.
  9. 9. The method according to claim 7, wherein in the second step, the pneumatic system (2) increases the air pressure in the micro-fluidic control chamber (131) by 2kPa relative to the atmospheric pressure.
  10. 10. The method according to claim 7, further comprising determining a pneumatic system (2) to adjust the air pressure variation value of the micro-fluid regulating chamber (131) according to a preset pulling amplitude of the transmission layer (12).

Description

Microfluidic-mediated static and dynamic collaborative cell culture chip and application thereof Technical Field The invention relates to the technical field of biology, in particular to a microfluidic-mediated static and dynamic collaborative cell culture chip and application thereof. Background The traditional technical path in the cell and tissue culture field can be summarized into three points, namely, one point is that a bearing medium is mainly a culture dish, a porous plate and the like; secondly, static two-dimensional culture is carried out in a culture mode, thirdly, the regulation and control mode is carried out by depending on active components such as growth factors in culture solution, and the like. The rise of micro-fluidic chip technology makes the traditional path change, namely the cross-boundary fusion of the two, become the core development trend of the field. The chip laboratory is a typical name of a microfluidic chip, and has the core advantages of being capable of accurately controlling fluid in a micron-sized space, integrating traditional laboratory functions such as chemical analysis and biological detection on a chip of only a few square centimeters, and simultaneously having a series of characteristics of less material consumption, micro reagent consumption, high reaction efficiency, excellent detection sensitivity, strong portability, simple and convenient operation, flexible combination of unit technologies and the like. Polydimethylsiloxane (PDMS) is the dominant fabrication material for such chips. The excellent light transmittance, gas permeability, biocompatibility and flexibility endow the material with unique advantages, and the material can be widely applied in the research fields of biochemistry, molecular biology and the like. By means of the PDMS chip, researchers can construct a composite culture microenvironment close to the in-vivo growth state of the cell tissue, can accurately regulate and control key mechanical parameters in the microenvironment, and meanwhile, the dynamic response process of the cell tissue generated by the change of the microenvironment parameters can be tracked and observed in real time. In the current related art system, many short plates have not been overcome, and the following two aspects are specifically shown: 1. The key regulation and control effects of biomechanical factors on cell tissue behaviors and functions are not considered by the traditional static two-dimensional culture system. The in-vivo real mechanical microenvironment cannot be re-carved, so that the in-vitro culture result and the in-vivo physiological state are obviously deviated, and the natural physiological characteristics of cells are difficult to be truly reflected. 2. In a simple dynamic culture mode or a static culture mode, stability and uniformity of microenvironment parameters are difficult to ensure, wherein the stability and uniformity of nutrient components such as growth factors in culture solution and the mechanical parameters such as fluid shear force are easy to fluctuate. The direct influence of the fluctuation is that researchers cannot efficiently and accurately distinguish which difference exists in regulating and controlling the physiological functions of cells under the two conditions of mechanical traction mechanical stimulation and static culture. The problem not only restricts the deep research of related regulation and control mechanisms, but also reduces the efficiency and result credibility of drug screening work. In summary, the technical problem that the scientific researchers in the field need to overcome is to develop a tissue chip capable of synchronously realizing static culture and mechanical traction state culture on the same chip platform. Disclosure of Invention In view of the analysis, the invention aims to provide a microfluidic mediated static and dynamic collaborative cell culture chip and application thereof, which are used for solving one of the problems that static culture is difficult to multiplex the real mechanical microenvironment in vivo, a chip device capable of synchronously implementing static and mechanical stretching dynamic biphase culture is lacking, and differential effects of stretching mechanical stimulation and static culture on cell physiological regulation are difficult to accurately define due to uneven microenvironment parameters in the prior art. The aim of the invention is mainly realized by the following technical scheme: The micro-fluidic mediated static and dynamic collaborative cell culture chip comprises a cell culture chip and a pneumatic system matched with the cell culture chip for use, wherein the cell culture chip comprises a culture layer, a transmission layer and a micro-fluidic bottom layer, the culture layer is provided with a culture chamber, the culture chamber comprises a static culture area and a dynamic culture area, and the micro-fluidic bottom layer is provided with a micro-fluid