CN116948823A - Microfluidic chip for heterogeneous cell culture and monitoring and preparation method thereof - Google Patents
Microfluidic chip for heterogeneous cell culture and monitoring and preparation method thereof Download PDFInfo
- Publication number
- CN116948823A CN116948823A CN202310934009.9A CN202310934009A CN116948823A CN 116948823 A CN116948823 A CN 116948823A CN 202310934009 A CN202310934009 A CN 202310934009A CN 116948823 A CN116948823 A CN 116948823A
- Authority
- CN
- China
- Prior art keywords
- channel
- gel
- cell
- tumor
- channels
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000012544 monitoring process Methods 0.000 title claims abstract description 24
- 238000004113 cell culture Methods 0.000 title claims abstract description 19
- 238000002360 preparation method Methods 0.000 title abstract description 8
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 47
- 210000004881 tumor cell Anatomy 0.000 claims abstract description 39
- 239000004205 dimethyl polysiloxane Substances 0.000 claims abstract description 30
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 claims abstract description 30
- 230000003993 interaction Effects 0.000 claims abstract description 22
- 230000010412 perfusion Effects 0.000 claims abstract description 21
- 239000003814 drug Substances 0.000 claims abstract description 18
- 238000011282 treatment Methods 0.000 claims abstract description 17
- 239000000758 substrate Substances 0.000 claims abstract description 13
- 239000011664 nicotinic acid Substances 0.000 claims abstract description 8
- 239000011521 glass Substances 0.000 claims abstract description 7
- 238000011161 development Methods 0.000 claims abstract description 6
- -1 Polydimethylsiloxane Polymers 0.000 claims abstract description 5
- 239000002131 composite material Substances 0.000 claims abstract description 3
- 210000004027 cell Anatomy 0.000 claims description 38
- 238000000034 method Methods 0.000 claims description 20
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 claims description 17
- 229910052710 silicon Inorganic materials 0.000 claims description 17
- 239000010703 silicon Substances 0.000 claims description 17
- 239000007788 liquid Substances 0.000 claims description 13
- 239000000523 sample Substances 0.000 claims description 13
- 210000001519 tissue Anatomy 0.000 claims description 10
- 230000008859 change Effects 0.000 claims description 9
- 230000008569 process Effects 0.000 claims description 9
- 238000003501 co-culture Methods 0.000 claims description 8
- 238000011049 filling Methods 0.000 claims description 8
- 230000008614 cellular interaction Effects 0.000 claims description 7
- 239000001963 growth medium Substances 0.000 claims description 7
- 229920002120 photoresistant polymer Polymers 0.000 claims description 7
- 239000006285 cell suspension Substances 0.000 claims description 6
- 239000012530 fluid Substances 0.000 claims description 6
- 238000010438 heat treatment Methods 0.000 claims description 6
- 238000006243 chemical reaction Methods 0.000 claims description 5
- 230000018109 developmental process Effects 0.000 claims description 5
- 238000003384 imaging method Methods 0.000 claims description 5
- 239000000126 substance Substances 0.000 claims description 5
- 238000009792 diffusion process Methods 0.000 claims description 4
- 239000007850 fluorescent dye Substances 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 235000015097 nutrients Nutrition 0.000 claims description 4
- 238000009832 plasma treatment Methods 0.000 claims description 4
- 238000002444 silanisation Methods 0.000 claims description 4
- 238000004528 spin coating Methods 0.000 claims description 4
- 230000007774 longterm Effects 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 230000002503 metabolic effect Effects 0.000 claims description 3
- 238000001259 photo etching Methods 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 claims description 3
- 238000004080 punching Methods 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 102000004127 Cytokines Human genes 0.000 claims description 2
- 108090000695 Cytokines Proteins 0.000 claims description 2
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 claims description 2
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 claims description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 2
- 230000010261 cell growth Effects 0.000 claims description 2
- 230000008878 coupling Effects 0.000 claims description 2
- 238000010168 coupling process Methods 0.000 claims description 2
- 238000005859 coupling reaction Methods 0.000 claims description 2
- 238000004132 cross linking Methods 0.000 claims description 2
- 238000005520 cutting process Methods 0.000 claims description 2
- 210000002744 extracellular matrix Anatomy 0.000 claims description 2
- 238000013427 histology analysis Methods 0.000 claims description 2
- 229920001477 hydrophilic polymer Polymers 0.000 claims description 2
- 238000003709 image segmentation Methods 0.000 claims description 2
- 108020004999 messenger RNA Proteins 0.000 claims description 2
- 239000002207 metabolite Substances 0.000 claims description 2
- 239000001301 oxygen Substances 0.000 claims description 2
- 229910052760 oxygen Inorganic materials 0.000 claims description 2
- 102000004169 proteins and genes Human genes 0.000 claims description 2
- 230000004044 response Effects 0.000 claims description 2
- 229940126585 therapeutic drug Drugs 0.000 claims description 2
- 230000001066 destructive effect Effects 0.000 claims 1
- 239000002245 particle Substances 0.000 claims 1
- 229940079593 drug Drugs 0.000 abstract description 14
- 230000035945 sensitivity Effects 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 5
- 238000011160 research Methods 0.000 abstract description 4
- 206010059866 Drug resistance Diseases 0.000 abstract description 3
- 239000002547 new drug Substances 0.000 abstract description 3
- 238000012546 transfer Methods 0.000 abstract description 3
- 206010061818 Disease progression Diseases 0.000 abstract description 2
- 230000005750 disease progression Effects 0.000 abstract description 2
- 238000009509 drug development Methods 0.000 abstract description 2
- 238000000338 in vitro Methods 0.000 abstract description 2
- 239000000499 gel Substances 0.000 description 26
- 210000002220 organoid Anatomy 0.000 description 8
- 238000004393 prognosis Methods 0.000 description 7
- 230000002792 vascular Effects 0.000 description 6
- 239000002202 Polyethylene glycol Substances 0.000 description 5
- 230000005012 migration Effects 0.000 description 5
- 238000013508 migration Methods 0.000 description 5
- 229920001223 polyethylene glycol Polymers 0.000 description 5
- 206010027476 Metastases Diseases 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- 238000013461 design Methods 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000007877 drug screening Methods 0.000 description 4
- 230000009401 metastasis Effects 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 238000011156 evaluation Methods 0.000 description 3
- 230000002269 spontaneous effect Effects 0.000 description 3
- 230000002123 temporal effect Effects 0.000 description 3
- IYMAXBFPHPZYIK-BQBZGAKWSA-N Arg-Gly-Asp Chemical compound NC(N)=NCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O IYMAXBFPHPZYIK-BQBZGAKWSA-N 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000001574 biopsy Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- IJOOHPMOJXWVHK-UHFFFAOYSA-N chlorotrimethylsilane Chemical compound C[Si](C)(C)Cl IJOOHPMOJXWVHK-UHFFFAOYSA-N 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 210000002889 endothelial cell Anatomy 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000000017 hydrogel Substances 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 210000000130 stem cell Anatomy 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 230000008685 targeting Effects 0.000 description 2
- 238000011269 treatment regimen Methods 0.000 description 2
- 230000004614 tumor growth Effects 0.000 description 2
- 102000008946 Fibrinogen Human genes 0.000 description 1
- 108010049003 Fibrinogen Proteins 0.000 description 1
- 206010066901 Treatment failure Diseases 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 108010072041 arginyl-glycyl-aspartic acid Proteins 0.000 description 1
- 230000006399 behavior Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003592 biomimetic effect Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000005929 chemotherapeutic response Effects 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 229940012952 fibrinogen Drugs 0.000 description 1
- 230000004907 flux Effects 0.000 description 1
- 230000014509 gene expression Effects 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 230000005660 hydrophilic surface Effects 0.000 description 1
- 230000005661 hydrophobic surface Effects 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 230000008611 intercellular interaction Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 206010061289 metastatic neoplasm Diseases 0.000 description 1
- 210000004088 microvessel Anatomy 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 238000000059 patterning Methods 0.000 description 1
- 238000000206 photolithography Methods 0.000 description 1
- 239000012985 polymerization agent Substances 0.000 description 1
- 230000002980 postoperative effect Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000000306 recurrent effect Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000001338 self-assembly Methods 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000005051 trimethylchlorosilane Substances 0.000 description 1
- 230000005748 tumor development Effects 0.000 description 1
- 230000006426 vascular sprouting Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/08—Bioreactors or fermenters specially adapted for specific uses for producing artificial tissue or for ex-vivo cultivation of tissue
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502707—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the manufacture of the container or its components
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/16—Microfluidic devices; Capillary tubes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/20—Material Coatings
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/14—Scaffolds; Matrices
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/10—Perfusion
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/08—Chemical, biochemical or biological means, e.g. plasma jet, co-culture
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
- C12M37/04—Seals
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/12—Specific details about manufacturing devices
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Sustainable Development (AREA)
- Clinical Laboratory Science (AREA)
- Molecular Biology (AREA)
- Dispersion Chemistry (AREA)
- Immunology (AREA)
- Cell Biology (AREA)
- Analytical Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Physiology (AREA)
- Hematology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention belongs to the technical field of microfluidic chips, and particularly relates to a microfluidic chip for heterogeneous cell culture and monitoring and a preparation method thereof. The microfluidic chip consists of a Polydimethylsiloxane (PDMS) substrate and a glass sheet, wherein a composite structure formed by combining a plurality of functional units is designed on the PDMS substrate, and each functional unit is provided with a perfusion channel, a gel channel, a volcanic capillary passive valve and a tumor cell channel. The microfluidic chip can be used for long-time and high-activity culture of heterogeneous tumor cells in an in-vitro three-dimensional bionic environment, realizes real-time dynamic observation of multi-cell interaction, monitors differences of different subtype tumor cells in aspects of functional characteristics, drug sensitivity, occurrence development speed, transfer potential and the like, and provides a novel high-flux bionic platform for personalized treatment, tumor drug resistance research, disease progression prediction and new drug development.
Description
Technical Field
The invention belongs to the technical field of microfluidic chips, and particularly relates to a microfluidic chip for heterogeneous cell culture and monitoring and a preparation method thereof.
Background
Cell heterogeneity (heterogenesis) is a phenomenon that is widely present in cellular systems, and is closely related to stem cell differentiation and the occurrence and prognosis of diseases such as cancer. The genome of the cell samples obtained from the same tissue may be identical between cells, but there are significant differences in morphology, behaviours, gene expression, and the like. However, traditional cell population-based research methods typically only average a population of cells, ignoring cell-to-cell variability, which presents a significant challenge for disease mechanism exploration and accurate treatment.
Tumor heterogeneity refers to the change in molecular biology or gene between daughter cells and mother cells in the course of tumor development due to multiple proliferation, which results in differences in tumor growth rate, invasion capacity, drug sensitivity, postoperative survival time, and patient prognosis [1] . Tumor heterogeneity includes spatial and temporal heterogeneity, the former referring to differences in different regions of the same tumor, and the latter referring to differences between primary and recurrent, secondary metastatic tumors. Tumor heterogeneity is considered to be one of the important causes of high mortality and drug resistance, and the higher the degree of heterogeneity, the worse the drug treatment effect and patient prognosis [2] . Therefore, understanding tumor heterogeneity is of great value in clinical diagnosis, personalized treatment, prognostic monitoring, and the like of tumors.
However, current clinical tumor diagnosis relies on tissue biopsy techniques for small-scale tumor feature assessment, and cannot represent the overall tumor of the patient; meanwhile, when the tumor is sampled after the operation, only malignant parts in the tumor are usually focused, the detection of invasive and heterogeneous areas is neglected, and the sensitivity and monitoring prognosis of the tumor to different treatment means cannot be accurately estimated in time. Furthermore, the temporal heterogeneity of tumors requires observation and detection at different stages of development, whereas tissue biopsies belong to invasive examinations and cannot be performed multiple times.
Disclosure of Invention
The invention aims to overcome the defects of the prior art, provides a microfluidic chip for heterogeneous cell culture and interaction monitoring and a preparation method thereof, and provides a novel high-flux bionic platform for personalized accurate treatment, drug sensitivity, disease progression monitoring and new drug development.
The invention adopts a microfluidic chip technology to establish a three-dimensional bionic co-culture system in vitro, carries out long-time, high-activity and high-flux culture on heterogeneous tumor cells, realizes real-time dynamic observation of multi-cell interaction, and monitors the differences of different subtype tumor cells in the aspects of functional characteristics, drug sensitivity, development speed, transfer potential and the like.
The microfluidic chip, also called chip laboratory, is a technology for controlling fluid in micrometer scale space, and can integrate basic operation units of sample preparation, reaction, separation, detection, cell culture, separation, cracking and the like in the fields of chemistry, biology and the like onto one chip.
The microfluidic chip for heterogeneous cell culture and interaction monitoring provided by the invention adopts Polydimethylsiloxane (PDMS) as a substrate material, and has good optical characteristics, insulativity, thermal stability and biocompatibility. And (3) mixing the prepolymer and the polymerizer (8-15) with the formula 1, pouring the mixture on a photoetching patterning template, solidifying the mixture to form a PDMS substrate, and bonding the PDMS substrate with a glass sheet through plasma treatment to prepare the microfluidic chip suitable for heterogeneous cell culture and interaction monitoring. The microfluidic chip has the characteristic of multi-unit integration, and can be used for researching the heterogeneity of cell populations in tumor tissues of a patient source by increasing the unit number, and is used for carrying out drug screening and personalized treatment.
The invention provides a micro-fluidic chip for heterogeneous cell culture and interaction monitoring, which consists of a PDMS substrate and a glass sheet which are bonded; wherein, the PDMS substrate is provided with a plurality of functional units which are combined in a certain form to form a composite structure, and each functional unit is provided with: perfusion channels, gel channels, volcanic capillary passive valves, and tumor cell channels; see fig. 1. Wherein:
the perfusion channel is used for perfusing various types of culture liquids, such as culture medium, cell suspension and drug solution; the structure of the perfusion channel is as follows: the structure of the arc-shaped (such as a semicircle-shaped) pipe cavity is taken as a unit, N units are sequentially communicated to form a continuous closed channel, and the same fluid environment is provided for each functional unit; the filling channel is provided with two sample inlets for connecting a filling system and filling various types of fluids into the filling channel;
the gel channel is used for pouring gel with various viscosities and gel mixed liquid formed by cells; the gel is formed by crosslinking hydrophilic polymer chains, can form a three-dimensional network structure, provides a three-dimensional support for cell growth, simultaneously provides support for diffusion and exchange of oxygen, nutrient substances and metabolic wastes, and plays a role of extracellular matrix; the number of the gel channels is N, and independent sample inlets are formed at two ends of each gel channel; the gel channel main body is of an arc-shaped tube cavity structure, one side of the gel channel main body is clung to the perfusion channel, and the other side of the gel channel main body is clung to the tumor cell channel; a plurality of arc trapezoid columns are uniformly distributed on two sides of the inner wall of the arc-shaped tube cavity to form an arc trapezoid column array, volcanic capillary passive valves are formed between two adjacent arc trapezoid columns, the 'contact angle hysteresis phenomenon' is utilized to change the local capillary force direction, the liquid flow is controlled, gel-like substances are further effectively limited in a gel channel, and long-term independent cell culture is realized; compared with the traditional linear passive valve, the volcanic capillary passive valve has stronger stability, can adapt to the pouring of hydrogels with different viscosities and different flow rates in a larger range, and greatly reduces the rupture of the capillary passive valve caused by the blocking of hydrogel injection or the too fast flow rate in the traditional design; meanwhile, the volcanic capillary passive valve can be applied to observation and research of cell interface biology, and the application range of the passive valve is widened;
the tumor cell channels are respectively arranged at the inner sides of the N gel channels, one end of the tumor cell channels is of a fan-shaped structure, the tumor cell channels are separated from the gel channels by taking the volcanic capillary passive valve as a boundary, and the other end of the tumor cell channels is a sample inlet for injecting cell suspension; the tumor cell channels of the functional units are independent of each other and do not affect each other, and each tumor cell channel is an independent reaction tank.
In the present invention, the number of integratable functional units N per chip is at least 3, typically 3-8, or more.
In the invention, a plurality of functional units share the same perfusion channel, thereby facilitating rapid high-flux perfusion culture and reducing repeated operation.
In the invention, the tumor cell channels of the functional units are independent of each other and do not affect each other, and each tumor cell channel is an independent reaction tank.
In the invention, each unit tumor channel is in a fan-shaped structure, the radius of the fan is 50-4000 mu m, the radian is 30-300 DEG, and the height is 10-200 mu m.
In the invention, each unit pouring channel and each gel channel are of an arc-shaped tube cavity structure, the width of the channel is 50-4000 mu m, and the height of the channel is 10-200 mu m.
In the invention, the length of the upper bottom of the arc trapezoid column is 10-850 mu m, the length of the lower bottom is 20-1000 mu m, the length of the lower bottom is longer than the length of the upper bottom, and the width between the upper bottom and the lower bottom is 10-200 mu m; preferably 300-500 μm for the upper base, 350-600 μm for the lower base, and 40-65 μm for the width between the upper base and the lower base. More preferably, the upper substrate is 400 μm, the lower substrate is 550 μm, and the width between the upper and lower substrates is 55 μm.
In the invention, the upper bottom spacing of the volcanic capillary passive valve formed between the arc trapezoid columns is 5-100 mu m, the lower bottom spacing is 10-200 mu m, the lower bottom spacing is larger than the upper bottom spacing, and the arc angle alpha ranges from 90 degrees to 180 degrees. Preferably, the upper sole spacing is 20-50 μm, the lower sole spacing is 80-130 μm, and the arc angle is 90 DEG to 124 deg. More preferably, the upper sole spacing is 30 μm, the lower sole spacing is 110 μm, and the arc angle is 108 °.
The preparation method of the multicellular heterogeneity interaction microfluidic chip provided by the invention comprises the following specific steps:
(1) Manufacturing a micro-fluidic chip structure pattern silicon wafer template by a photoetching method, wherein the micro-fluidic chip structure pattern silicon wafer template comprises the steps of spin coating photoresist, soft baking, ultraviolet exposure, post baking, development, hardening, silanization treatment and the like;
(2) Mixing and stirring Polydimethylsiloxane (PDMS) prepolymer and a polymerizer in a mass ratio of (8-15): 1, and centrifuging to remove bubbles;
(3) Pouring the mixed and centrifuged PDMS on a patterned silicon wafer template, vacuumizing to remove bubbles, transferring to an 80 ℃ oven, and heating until the PDMS is solidified;
(4) Cutting and taking out the cured PDMS chip from the template, and punching a sample hole according to the requirement;
(5) The PDMS chip and the glass sheet were tightly attached by plasma treatment or by using a jig to ensure that the liquid did not leak.
The invention relates to a multicellular heterogeneity interaction micro-fluidic chip, which comprises the following operation procedures:
(1) Placing the prepared microfluidic chip in an oven at 80 ℃ for 24 hours, and recovering the hydrophobicity in the channel;
(2) Before the chip is used, ultraviolet irradiation is carried out for 1 hour for sterilization;
(3) Injecting gel solution into the gel channel from a gel channel sample inlet, and pouring culture liquid into the pouring channel after gel is solidified;
(4) The cell suspension is introduced into the tumor cell channel from a tumor cell channel sample inlet, and polyethylene glycol (PEG) and arginyl-glycyl-aspartic acid (RGD) with different proportions are modified in the channel in advance according to the requirement so as to change the adhesiveness of the channel substrate.
The multicellular heterogeneity interaction microfluidic chip provided by the invention can be used for heterogeneous cell culture and interaction monitoring, and is concretely as follows:
(1) The invention can be used for three-dimensional bionic co-culture among different cells, and the channels can be relatively independent at the beginning, but small molecules such as nutrient substances, cytokines and the like are communicated; fresh culture medium can be continuously provided in the continuous perfusion culture process, a stable environment is provided for long-time culture of cells, and meanwhile, culture medium liquid can be continuously collected for downstream histology analysis and the like;
(2) The invention is perfectly adapted to a high-resolution microscopic imaging system, and can realize rapid dynamic and high-resolution cell-cell interaction monitoring; the method is matched with a living cell culture system (such as okolab), multi-channel and multi-marked fluorescence or bright field comprehensive scanning is carried out on cells and subcellular even macromolecular layers, six-dimensional (x, y, z, time, multi-wavelength and multi-position) real-time recording and observation of a dynamic process are realized, and comprehensive cell interaction information is obtained;
(3) By combining continuous perfusion and real-time dynamic imaging of the chip, the invention can uniformly perfuse interference factors or therapeutic drugs to heterogeneous cells, and monitor and evaluate the responses of different subtype cells to the same intervention object for a long time; the dynamic changes of cells and tissues are effectively and accurately captured, the identification, positioning, tracking, image segmentation and the like of different cells are realized, and the interaction among heterogeneous cells is further quantitatively analyzed and compared;
(4) The invention can integrate the sensor to monitor the cell dynamically and continuously without intervention or destruction; by coupling the chip with the electric sensor, monitoring the overall impedance on the chip to reflect the impedance value change in the cell interaction process, and capturing the heterogeneity of the cell interaction; the fluorescent biosensor can also be coupled, such as fluorescent dye, fluorescent probe, nano particles and the like, and can monitor the luminous intensity, reflection index and incident/reflected light angle change of various proteins, metabolites and mRNA to realize dynamic monitoring of molecular level.
The design of the microfluidic chip for heterogeneous cell culture and interaction monitoring has the following characteristics:
(1) A stable volcanic capillary passive valve can be formed between arc trapezoid columns designed on the chip, and the 'contact angle hysteresis phenomenon' is utilized to change the local capillary force direction, so that the liquid flow is controlled, and the stable construction of a cell interface in the chip and the long-term independent cell culture are realized;
(2) The arc angle alpha of the volcanic capillary passive valve in the design has a good effect in the range from 90 degrees to 124 degrees, can effectively disperse the liquid pressure in the gel channel, satisfies the requirements that gel with various viscosities in a larger range can smoothly fill the whole gel channel at a uniform speed, and ensures that the capillary passive valve is not broken;
(3) The low adhesion pretreatment of the tumor channel can meet the requirement of spontaneous balling on a tumor cell chip, and provides a more bionic three-dimensional tumor tissue structure;
(4) The communicated perfusion channels facilitate the perfusion of the culture medium and the drug solution, and repeated operation is reduced;
(5) The microfluidic chip can be used for heterogeneous analysis of multicellular interaction, and the design of the functional units can be repeatedly integrated and combined, so that the microfluidic chip can meet the requirement of high flux more than the conventional microfluidic chip with a single structure.
The microfluidic chip designed by the invention has the following advantages:
(1) The compatibility is strong, the cost is low, the biocompatibility is good, and the operation is simple and convenient;
(2) The invention can realize the co-culture of different cells, organoids and tissues, is used for researching the interaction of the cells, the organoids and the tissues, and compared with other co-culture systems, the invention is closer to an in-vivo microenvironment, and has the advantages of dynamic real-time observation, good biocompatibility and small sample size;
(3) Functional units can be repeatedly added and arranged and combined to meet the requirement of cell heterogeneity analysis:
aiming at the tumor space heterogeneity, medicine screening can be carried out on a chip, so that the medicine strategy of different tumor areas is optimized, and the treatment targeting is enhanced;
aiming at the tumor time heterogeneity, tumor cells can be cultured on a chip for a long time with high activity, the whole process of tumor growth, diffusion and metastasis can be dynamically observed in real time, the tumor progress speed and the metastasis potential can be predicted, and doctors can be assisted to develop more accurate prognosis evaluation and formulate corresponding intervention measures;
(4) The chip can be used for carrying out quantification and comparison of migration capacity of different cell subtypes and induction of vascular sprouting capacity in tumors, can effectively distinguish tumor cells with strong migration capacity and weak migration capacity, and separates cell subpopulations with specific migration capacity from heterogeneous tumor cell populations, thereby providing possibility for personalized medicine and drug screening;
(5) The chip can be used for researching immune scores of cancer treatment, obtains more detailed chemo-therapeutic response trend through co-culture with immune cells, provides a classification standard based on tumor immune microenvironment interaction mode, and provides a theoretical basis for guiding individual treatment of cancer;
(6) The volcanic capillary passive valve formed by optimization is stable and reliable, has good repeatability and is suitable for gels with various viscosities.
Drawings
Fig. 1 is a schematic diagram of a structure of a biomimetic microfluidic chip for heterogeneous cell culture and interaction monitoring, which are formed by combining six functional units.
Fig. 2 shows the structure of each part in a single functional unit.
Fig. 3 shows spontaneous tumor cell formation of tumor spheres on a low adhesion surface treated microfluidic chip.
Reference numerals in the drawings: 1 is a perfusion channel; 2 is a gel channel; 3 is an arc trapezoid column; 4 is tumor cell channel; and 5 is a volcanic capillary passive valve.
Detailed Description
The invention is further described below with reference to examples and figures.
Example 1 a microfluidic chip for interaction of a tumor organoid and a self-assembled vascular network comprising six functional units (see fig. 1), each designed with perfusion channels, gel channels, arced trapezoidal columns, tumor cell channels. Wherein, the mixed solution of fibrinogen (3 mg/mL) and endothelial cells is introduced into the gel channel to induce the self-assembly of the gel channel to form a microvascular network. Then modifying polyethylene glycol PEG 3h (0.5 mg/mL) in the cell channel to form hydrophobic surface, introducing tumor cell suspension (cell density: 1.5X10) at different positions of tumor tissue of the same patient 6 and/mL), after 24h the tumor cells spontaneously formed spherical tumor organoids, after which the spheres were continuously regularized, with spheres being most regular around 150h, 240 μm in diameter, and compact (fig. 3). The interaction between the tumor organoids and the vascular network is monitored in real time, the migration capacity and the angiogenesis induction capacity of the tumor organoids of different units are compared and analyzed, then medicines are added for intervention, and the influence of the medicines on tumor cells of different subtypes is evaluated.
The preparation method of the microfluidic chip for interaction between the heterogeneous tumor organoid and the self-assembled vascular network provided by the invention comprises the following specific steps:
(1) Drawing and designing a micro-channel structure of a chip by adopting AutoCAD software, preparing a silicon template with a designed micro-pattern by using a photolithography method, wherein the steps comprise spin-coating photoresist, soft baking, ultraviolet exposure, post baking, developing, hardening and silanization treatment; the specific operation is as follows: uniformly spin-coating a photoresist SU-8 2050 on a silicon wafer (1700 rpm), and then sequentially placing the silicon wafer on a 65 ℃ heating table for 5min and a 95 ℃ heating table for 25min to finish a soft baking step; after the silicon wafer is cooled, placing a film with a designed pattern above the photoresist, and exposing the film to ultraviolet light for 30s; then placing the silicon wafer in a 65 ℃ heating table for 5min and a 95 ℃ heating table for 10min in sequence to finish the post-baking step; then placing the silicon wafer into a developing solution for developing for 9min; clear patterns formed by solidification of photoresist are visible on the developed silicon wafer, and then the silicon wafer is placed on a heat table at 150 ℃ for hardening for 30min; finally, carrying out silanization treatment on the surfaces of the silicon wafer and the photoresist by using trimethylchlorosilane;
(2) Mixing and stirring Polydimethylsiloxane (PDMS) prepolymer and a polymerization agent in a mass ratio of 10:1, and centrifuging to remove bubbles;
(3) Pouring the mixed and centrifuged PDMS on a silicon plate, vacuumizing by a vacuum pump to remove bubbles, and placing the PDMS in an oven at 80 ℃ for 1h to solidify the PDMS;
(4) Taking down the solidified and cooled PDMS chip from the silicon template, and punching a sample injection hole with the diameter of 2mm by a puncher;
(5) Bonding a hydrophilic surface PDMS chip and a glass sheet after plasma treatment, and then placing the whole microfluidic chip in an oven at 80 ℃ for 24 hours to recover the hydrophobicity in the channel;
(6) Before the chip is used, ultraviolet irradiation is carried out for 1h for sterilization.
In general, tumor spatial and temporal heterogeneity, multiple sampling at multiple points is often required clinically, and then molecular biological analysis is performed on individual samples, respectively, with high cost, long time consumption, large errors and unfavorable patient recovery in the whole process. The micro-fluidic chip designed by the invention is used for culturing, detecting and analyzing the multicellular. Each unit is a single reaction chamber, and can perform long-time detection analysis on tumor cells at different parts of the same patient. The tumor heterogeneity is known to be an important cause of drug treatment failure, and the drug sensitivity of different subtype cells is different, so that drug screening can be carried out on heterogeneous tumor cells of a patient in each unit, an optimal drug strategy of different areas of the tumor is obtained, and the targeting of the drug is enhanced to realize accurate treatment. Secondly, the growth and diffusion speed of the tumor and the transfer potential thereof can be predicted by long-time culture of heterogeneous tumor cells, which is beneficial to prognosis evaluation of patients, timely adjustment of treatment strategies and the like. In addition, the tumor heterogeneity research can guide the development of new drugs, and the targeted drugs and corresponding treatment schemes are developed according to the different molecular characteristics and metabolic modes of different subtype tumor cells, so that personalized layered treatment is realized.
The micro-fluidic chip has good biocompatibility, is easy to operate and observe, and is matched with most imaging systems, including an optical microscope, a split microscope, a laser confocal microscope, a two-photon microscope, a total internal reflection fluorescence microscope and the like. Compared with the traditional cell co-culture Transwell method, the method can better simulate the three-dimensional environment and fluid state in vivo. The chip of the invention can induce endothelial cells to self-assemble to form perfusable vascular networks with different diameters, and dynamically observe the generation process of the vascular networks and the interaction between tumor organoids and the vascular networks in real time. In addition, the spontaneous formation of tumor spheres by PEG modification of the channel is simpler and reduces the damage that may be caused to the tumor sphere during metastasis, compared to manual metastasis of the tumor sphere to the channel. The invention has wide application prospect in the establishment of qualitative grading of tumor, guiding personalized treatment strategy, drug screening evaluation and early prediction of patient prognosis.
Reference to the literature
[1]De Sousa EMF,Vermeulen L,Fessler E,et al.Cancer heterogeneity--a multifaceted view[J].EMBO Rep,2013,14(8):686-695.
[2]Burrell RA,Swanton C.Tumour heterogeneity and the evolution of polyclonal drug resistance[J].Mol Oncol,2014,8(6):1095-1111。
Claims (8)
1. The microfluidic chip for heterogeneous cell culture and monitoring is characterized by being formed by bonding a PDMS substrate and a glass sheet; wherein, the PDMS substrate is provided with a composite structure formed by combining a plurality of functional units, and each functional unit is provided with: perfusion channels, gel channels, volcanic capillary passive valves, and tumor cell channels; wherein:
the perfusion channel is used for perfusing culture liquid, including culture medium, cell suspension or medicine solution; the structure of the perfusion channel is as follows: the structure of the arc-shaped pipe cavity is taken as a unit, N units are sequentially communicated to form a continuous closed channel, and the same fluid environment is provided for each functional unit; the filling channel is provided with two sample inlets for connecting a filling system and filling various types of fluids into the filling channel;
the gel channel is used for pouring gel with various viscosities and gel mixed liquid formed by cells; the gel is formed by crosslinking hydrophilic polymer chains and is used for forming a three-dimensional network structure, providing a three-dimensional scaffold for cell growth, and simultaneously providing support for diffusion and exchange of oxygen, nutrients and metabolic wastes, and playing a role of extracellular matrix; the number of the gel channels is N, and independent sample inlets are formed at the two ends of each gel channel and are used for connecting a perfusion system; the gel channel main body is of an arc-shaped tube cavity structure, one side of the gel channel main body is clung to the perfusion channel, and the other side of the gel channel main body is clung to the tumor cell channel; a plurality of arc trapezoid columns are uniformly distributed on two sides of the inner wall of the arc-shaped lumen in the middle of each gel channel unit to form an arc trapezoid column array, volcanic capillary passive valves are formed between two adjacent arc trapezoid columns, the 'contact angle hysteresis phenomenon' is utilized to change the local capillary force direction, the liquid flow is controlled, gel-like substances are further effectively limited in the gel channels, and long-term independent cell culture is realized;
the tumor cell channels are respectively arranged at the inner sides of the N gel channels, the ends of the tumor cell channels are in a fan-shaped structure, the tumor cell channels are separated from the gel channels by taking the volcanic capillary passive valve as a boundary, and the ends of the tumor cell channels are provided with sample inlets for injecting cell suspension; the tumor cell channels of the functional units are independent of each other and do not affect each other, and each tumor cell channel is an independent reaction tank.
2. The microfluidic chip of claim 1, wherein N is at least 3.
3. The microfluidic chip according to claim 1, wherein the tumor channel has a fan-shaped structure, a fan-shaped radius of 50-4000 μm, an arc of 30 ° -300 °, and a height of 10-200 μm.
4. The microfluidic chip according to claim 1, wherein the perfusion channel and the gel channel are of an arc-shaped tube cavity structure, and the channel width is 50-4000 [ mu ] m, and the channel height is 10-200 [ mu ] m.
5. The microfluidic chip according to claim 1, wherein the arc trapezoid column has an upper bottom length of 10-850 μm, a lower bottom length of 20-1000 μm, a lower bottom length greater than the upper bottom length, and a width between the upper bottom and the lower bottom of 10-200 μm.
6. The microfluidic chip according to claim 1, wherein the volcanic capillary passive valve formed between the arched trapezoidal columns has an upper bottom spacing of 5-100 μm and a lower bottom spacing of 10-200 μm, the lower bottom spacing being larger than the upper bottom spacing, and the arc angle α is 90 ° -180 °.
7. A method for manufacturing a microfluidic chip according to any one of claims 1 to 6, comprising the following specific steps:
(1) Manufacturing a micro-fluidic chip structure pattern silicon wafer template by using a photoetching method, wherein the micro-fluidic chip structure pattern silicon wafer template comprises spin-coating photoresist, soft baking, ultraviolet exposure, post baking, development, hardening and silanization treatment steps;
(2) Mixing and stirring Polydimethylsiloxane (PDMS) prepolymer and a polymerizer in a mass ratio of (8-15): 1, and centrifuging to remove bubbles;
(3) Pouring the mixed and centrifuged PDMS on a silicon wafer template, vacuumizing to remove bubbles, transferring to an oven at 80 ℃ for heating until the PDMS is solidified;
(4) Cutting and taking out the cured PDMS chip from the template, and punching a sample hole according to the requirement;
(5) The PDMS chip and the glass sheet were tightly attached by plasma treatment or by using a jig to ensure that the liquid did not leak.
8. The microfluidic chip according to any one of claims 1 to 6 for heterogeneous cell culture and interaction monitoring, comprising:
(1) The three-dimensional bionic co-culture method is used for three-dimensional bionic co-culture among different cells, and all channels are relatively independent at the beginning, but nutrient substances and cytokines are communicated; fresh culture medium can be continuously provided in the continuous perfusion culture process, a stable environment is provided for long-time culture of cells, and meanwhile, culture medium liquid can be continuously collected for downstream histology analysis;
(2) The method is suitable for a high-resolution microscopic imaging system, and can realize rapid dynamic and high-resolution cell interaction monitoring; the living cell culture system is matched to carry out multi-channel and multi-marked fluorescence or bright field comprehensive scanning on cells and subcellular even macromolecular layers, thereby realizing six-dimensional, i.e. x, y, z, time, multi-wavelength and multi-position real-time recording and observation of a dynamic process and obtaining comprehensive cell interaction information;
(3) Combining continuous perfusion and real-time dynamic imaging of the chip, uniformly perfusing interference factors or therapeutic drugs to heterogeneous cells, and monitoring and evaluating the responses of different subtype cells to the same intervention object for a long time; the dynamic changes of cells and tissues are effectively and accurately captured, the identification, positioning, tracking and image segmentation of different cells are realized, and the interaction among heterogeneous cells is further quantitatively analyzed and compared;
(4) The integrated sensor is used for carrying out non-invasive and non-destructive dynamic continuous monitoring on the cells; by coupling the chip with the electric sensor, monitoring the overall impedance on the chip to reflect the impedance value change in the cell interaction process, and capturing the heterogeneity of the cell interaction; or coupled fluorescent biosensor, including fluorescent dye, fluorescent probe and nanometer particle, to monitor the luminous intensity, reflection index and incident/reflected light angle change of various proteins, metabolites and mRNA, to realize dynamic monitoring of molecular level.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310934009.9A CN116948823A (en) | 2023-07-27 | 2023-07-27 | Microfluidic chip for heterogeneous cell culture and monitoring and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310934009.9A CN116948823A (en) | 2023-07-27 | 2023-07-27 | Microfluidic chip for heterogeneous cell culture and monitoring and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116948823A true CN116948823A (en) | 2023-10-27 |
Family
ID=88461565
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310934009.9A Pending CN116948823A (en) | 2023-07-27 | 2023-07-27 | Microfluidic chip for heterogeneous cell culture and monitoring and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116948823A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113302276A (en) * | 2018-11-15 | 2021-08-24 | 弗拉斯沃克斯有限责任公司 | Dendritic cell generation apparatus and method |
| CN118314570A (en) * | 2024-06-11 | 2024-07-09 | 重庆医科大学绍兴柯桥医学检验技术研究中心 | White blood cell classification method and system for error correction of detection card |
-
2023
- 2023-07-27 CN CN202310934009.9A patent/CN116948823A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113302276A (en) * | 2018-11-15 | 2021-08-24 | 弗拉斯沃克斯有限责任公司 | Dendritic cell generation apparatus and method |
| CN118314570A (en) * | 2024-06-11 | 2024-07-09 | 重庆医科大学绍兴柯桥医学检验技术研究中心 | White blood cell classification method and system for error correction of detection card |
| CN118314570B (en) * | 2024-06-11 | 2024-08-06 | 重庆医科大学绍兴柯桥医学检验技术研究中心 | White blood cell classification method and system for error correction of detection card |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN116948823A (en) | Microfluidic chip for heterogeneous cell culture and monitoring and preparation method thereof | |
| US10725021B2 (en) | Muscle chips and methods of use thereof | |
| EP2335370B1 (en) | Organ-on-a-chip-device | |
| DE60003171T2 (en) | METHODS FOR MINIATURIZED CELL ARRANGEMENT AND CELL-BASED SCREENING DEVICE | |
| US11566224B2 (en) | Dendritic cell generator | |
| US8828332B2 (en) | Microfluidic capsule | |
| US9617520B2 (en) | Device and method of 3-dimensionally generating in vitro blood vessels | |
| Li et al. | Recent advances in an organ-on-a-chip: biomarker analysis and applications | |
| Pirlo et al. | Biochip/laser cell deposition system to assess polarized axonal growth from single neurons and neuron/glia pairs in microchannels with novel asymmetrical geometries | |
| CN112226363A (en) | Device and method for culturing high-flux organoid by utilizing microarray deep well | |
| CN114317272B (en) | Culture device for multicellular co-culture and cell culture method | |
| Xu et al. | Development of disposable PDMS micro cell culture analog devices with photopolymerizable hydrogel encapsulating living cells | |
| Johnson et al. | The applications and challenges of the development of in vitro tumor microenvironment chips | |
| US20230203417A1 (en) | Microfluidic device | |
| CN117721019A (en) | Vascularized organ chip and application thereof | |
| Saini et al. | Fabrication Method of a High-Density Co-Culture Tumor–Stroma Platform to Study Cancer Progression | |
| Saadin et al. | Enrichment of tumor-initiating breast cancer cells within a mammosphere-culture microdevice | |
| CN114874989B (en) | Method for capturing circulating tumor cells | |
| CN114989977A (en) | Tumor organoid chip for multi-cell interaction and drug screening and preparation method thereof | |
| CN214088510U (en) | Device structure for capturing circulating tumor cells | |
| Aydin et al. | Tumor-Microenvironment-on-Chip Platform for Assessing Drug Response in 3D Dynamic Culture | |
| JP5894732B2 (en) | Cell culture substrate evaluation method | |
| US11377634B2 (en) | 3D micro fabrication and screening | |
| US20240228950A1 (en) | Micropatterned 3d hydrogel microarray in fluidic channels for spheroid-in-gel culture | |
| CN116376694A (en) | Microfluidic chip for simulating tumor microenvironment and application method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |