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CN-122012239-A - Skin organoid chip based on multi-condition simulation detection

CN122012239ACN 122012239 ACN122012239 ACN 122012239ACN-122012239-A

Abstract

The invention belongs to the technical field of organoid culture, and provides a skin organoid chip based on multi-condition simulation detection. The invention comprises a gas flow layer, a culture layer, a porous membrane layer and a liquid flow layer which are sequentially laminated from top to bottom; the gas flow layer comprises a gas pump unit and a gas distribution structure connected in series with the gas pump unit, the gas pump unit is used for stabilizing gas flow, the gas distribution structure is used for distributing gas flow and buffering gas flow fluctuation, the culture layer comprises a plurality of culture holes communicated with the gas flow layer and a porous membrane layer, the porous membrane layer is used for supporting skin organoids and allowing small molecular substances to exchange, the liquid flow layer comprises a parallel liquid pump unit and a liquid distribution mechanism connected in series with the parallel liquid pump unit, the parallel liquid pump unit is used for stabilizing liquid flow, and the liquid distribution structure is used for distributing liquid and buffering liquid fluctuation. The invention aims to realize high-throughput culture and detection of skin organoids and meet the application in large-scale drug screening and toxicology evaluation.

Inventors

  • HONG MINHUA
  • ZHAO XIANGLONG
  • LIU JIN
  • WANG DIQING
  • WANG HUI
  • TANG RONG

Assignees

  • 杭州湃肽生化科技有限公司
  • 上海湃肽生物科技有限公司

Dates

Publication Date
20260512
Application Date
20260413

Claims (10)

  1. 1. The skin organoid chip based on multi-condition simulation detection is characterized by comprising a gas flow layer (1), a culture layer (2), a porous membrane layer (3) and a liquid flow layer (4) which are sequentially stacked from top to bottom, wherein the gas flow layer (1) comprises a gas pump unit and a gas distribution structure connected with the gas pump unit in series, the gas pump unit is used for stabilizing gas flow and buffering gas flow fluctuation, the culture layer (2) comprises a plurality of culture holes (21) communicated with the gas flow layer (1) and the porous membrane layer (3), the porous membrane layer (3) is used for supporting skin organoids and allowing small molecular substances to exchange, the liquid flow layer (4) comprises a parallel liquid pump unit and a liquid distribution mechanism connected with the parallel liquid pump unit in series, the liquid distribution structure is used for distributing liquid and buffering liquid fluctuation, the porous membrane layer (3) comprises a porous membrane (32) at the bottom of the culture hole (21) and a through hole matched with the porous membrane (32), the through hole (31) is arranged on the side wall, and the two ends of the piezoelectric membrane (31) are respectively connected with the piezoelectric membrane (32) in a compatible mode.
  2. 2. A skin organoid chip based on multi-condition simulation test according to claim 1, characterized in that the parallel liquid pump units of the liquid flow layer (4) are a plurality of liquid pump units connected in parallel, the liquid distribution structure comprises liquid channels, the number of liquid pump units is equal to the number of liquid channels of the liquid flow layer (4), each liquid pump unit has a corresponding liquid channel, and the liquid channels can be connected by using a multi-way valve with the number of outlets equal to the number of liquid channels.
  3. 3. A skin organoid chip based on multi-condition analog detection according to claim 2, wherein the liquid pump unit comprises micro metering pumps arranged at both ends of the liquid channel, the micro metering pumps being electrically connected to ensure that the volume of inflow and outflow liquid of the liquid channel is equal.
  4. 4. A skin organoid chip based on multi-condition simulation test according to claim 3, characterized in that the liquid channel comprises a liquid area (41) in the middle, the top of the liquid area (41) is in contact with the porous membrane layer (3) and passes through the bottom of the culture well (21), both ends of the liquid area (41) and the liquid channel are horizontally arranged at the liquid flow layer (4), and the height of the liquid area (41) is higher than both ends of the liquid channel.
  5. 5. A skin organoid chip based on multi-condition simulation detection according to claim 4, characterized in that the area of the liquid flow cross section of the liquid area (41) is larger than the area of the liquid flow cross sections at both ends of the liquid channel, the depth of the liquid area (41) is smaller than 1mm, the depth of the liquid area (41) is such that liquid flows within the liquid flow layer (4).
  6. 6. The skin organoid chip based on multi-condition simulation test according to claim 5, wherein a temperature control element (42) is arranged at the bottom of the liquid area (41), the temperature control element (42) corresponds to the culture hole (21), and a heat conducting silica gel layer is covered on the surface of the temperature control element (42) to transfer temperature to the skin organoid of the culture layer (2) through the liquid of the liquid area (41).
  7. 7. A skin organoid chip based on multi-condition simulation detection according to claim 1, characterized in that the gas flow layer (1) comprises gas channels (11), both ends of the gas channels (11) are provided with countersunk bends, the bottom of the gas channels (11) is in contact with the porous membrane layer (3) and passes the top of the culture wells (21).
  8. 8. A skin organoid chip based on multi-condition simulation detection according to claim 7, characterized in that the depth of the gas channel (11) is smaller than 1mm, the depth of the gas channel (11) being such that gas flows within the gas flow layer (1).
  9. 9. The skin organoid chip based on multi-condition simulation test according to claim 1, wherein the diameter of the porous membrane (32) at the bottom of each culture well (21) is consistent with that of the culture well (21), the edge extends to the bottom of the side wall of the culture well (21), the porous membrane (32) is made of at least one selected from polymethyl methacrylate, polydimethylsiloxane or polycarbonate, and the pore diameter of the porous membrane (32) is selected from the range of 0.4-5 μm.
  10. 10. The skin organoid chip of claim 1, wherein the piezoelectric element (31) is a fan-shaped piezoelectric ceramic sheet, and the piezoelectric ceramic sheet can be driven by voltage to generate expansion and contraction deformation, so as to drive the edge of the porous membrane to synchronously stretch, and the piezoelectric element can generate a voltage signal under the condition of being pressed and pulled.

Description

Skin organoid chip based on multi-condition simulation detection Technical Field The invention belongs to the technical field of organoid culture, and provides a skin organoid chip based on multi-condition simulation detection. Background The skin is the largest organ of human body and has multiple functions of barrier protection, immune defense, metabolic regulation and the like. With environmental pollution, aging of population, and rapid development of cosmetics and external medicines, skin-related diseases and medicine safety problems are increasingly prominent. In order to better study the physiological functions, disease mechanisms and toxicological effects of drugs and cosmetics of the skin, establishing an in vitro skin model is becoming an important means for replacing animal experiments. Compared with the traditional two-dimensional culture and animal models, the skin organoids have obvious advantages in simulating the structure and function of human skin, and have been widely applied to disease modeling, drug screening and personalized medicine research. However, the prior skin organoids still face the following technical bottlenecks in the application process, namely 1, the high-throughput culture and detection capability is insufficient, the traditional culture mode is mostly based on culture dishes or pore plates, the parallel culture and detection under large-scale and diversified conditions are difficult to realize, and the application of the skin organoids in drug screening and toxicology evaluation is severely limited. 2. The detection condition is single, namely the traditional organoid chip platform can only operate in a single environment or detection mode, and can not simultaneously meet the multi-condition detection requirements of medicine gradient, physical factor stimulation, multi-component microenvironment and the like. 3. The standardization and the compatibility are poor, the structure of part of chips is complex, the processing cost is high, the chips are difficult to be compatible with the existing automatic imaging and analysis system, and the popularization and the large-scale application are not facilitated. 4. The micro-environment control is insufficient, the skin organoids are highly sensitive to culture environments such as oxygen concentration, nutrient supply, mechanical stress and the like, but the existing culture systems have limited dynamic regulation and in-situ monitoring capability on the parameters, and influence the maturity of the organoids and the accuracy of functional evaluation. Disclosure of Invention The invention aims to realize high-throughput culture of skin organoids and simulated detection of multiple environmental conditions, and meet the requirements of application in large-scale drug screening and toxicology evaluation. Specifically, the multi-condition detection requirements such as oxygen concentration, nutrition supply, medicine gradient, mechanical stress, temperature gradient and other medicine gradients, physical factor stimulation, multi-component microenvironment and the like in the skin organoid culture environment can be adjusted, and the multi-detection condition detection of the skin organoid is realized. The skin organoid chip based on the multi-condition simulation detection comprises a gas flow layer, a culture layer, a porous membrane layer and a liquid flow layer which are sequentially stacked from top to bottom, wherein the gas flow layer comprises a gas pump unit and a gas distribution structure connected with the gas pump unit in series, the gas pump unit is used for stabilizing gas flow, the gas distribution structure is used for distributing gas flow and buffering gas flow fluctuation, the culture layer comprises a plurality of culture holes communicated with the gas flow layer and the porous membrane layer, the porous membrane layer is used for supporting skin organoids and allowing small molecular substances to exchange, the liquid flow layer comprises a parallel liquid pump unit and a liquid distribution mechanism connected with the parallel liquid pump unit in series, the parallel liquid pump unit is used for stabilizing liquid flow, and the liquid distribution structure is used for distributing liquid and buffering liquid fluctuation. The porous membrane layer also comprises a through hole matched with the porous membrane, the side wall of the through hole is provided with a piezoelectric element, and two ends of the piezoelectric element are respectively connected with the edge of the porous membrane and the through hole by adopting a biocompatible adhesive. The multilayer chip structure can replace one or more layers of chips according to the needs, is suitable for various application scenes, and the culture layer can be expanded according to the needs, so that the number of arrays of culture holes is increased, the function of high-throughput culture is realized, and the requirements of large-scale drug screening