3D Cell Culture Services

Our 3D Cell Culture Platform

Invitrocue’s expertise in 3D culture allows us to successfully recapitulate in vitro the normal and disease conditions of humans. Our 3D cell culture techniques using our 3D Cellulose Sponge scaffold, or non-scaffold-based techniques are more cost-effective and physiologically relevant compared to conventional 2D monolayer cultures.

By co-culturing and tri-culturing organ-specific cells with immune cells or endothelial cells, our 3D models are better able to recapitulate the microenvironment and functions of human organs, providing a more clinically relevant model for drug efficacy and toxicity testing.

2D cell culture

  • Lack of cell-to-cell and cell-ECM interaction
  • No gradients present
  • No drug resistance
  • Co-culture unable to establish a proper microenvironment
  • Poor clinical correlation

 

3D cell culture

  • Physiologic cell-to cell and ECM interaction
  • Drugs, oxygen, and nutrients diffuse in gradient
  • Drug resistance in vivo
  • Co-culture of multiple cell types mimic in vivo microenvironment
  • More reliable and better estimate of in vivo responses

 

Respiratory models

Invitrocue offers in vitro 3D respiratory models using air-liquid interphase culture for drug testing, inhalation toxicity, viral infections and inflammation studies.

Features:

  • Our 3D mucociliary tissue model consists of normal, human-derived nasal/bronchial epithelial cells
  • Our 3D structure consists of basal cells, goblet cells and ciliated cells with mucociliary clearance functions
  • Our model exhibits pseudostratified columnar epithelial morphology with high uniformity and reproducibility 
  • Our culture system preserves the physiological characteristics of nasal and bronchial epithelia

 

Inhalation Toxicity Test

Our inhalation toxicity test evaluates the toxic characteristics of inhalable materials, such as gases, volatile substances or aerosols/particulates. Acute inhalation toxicity data are used to satisfy hazard classification and labelling requirements, to estimate the toxicity of mixtures, and to assess human health and environmental risks.

Experimental design for drug testing using nasal/bronchial models

   Test Model   Nasal/Bronchial models
   Replicates   N=3 tissues per test condition
   Exposure Time   3-6 hours topical exposure to a predetermined 4-concentration range of test chemicals
   Assay Controls   Negative Control – Sterile DI H2O

   Positive Control – 14mg/ml Formaldehyde

   Endpoints   MTT, TEER, Marker staining, real-time PCR
   Data Delivery   Cell viability, permeability analysis,cell type and gene analysis

 

Anti-Viral Test

Our viral infected model for common cold (Rhinovirus) and flu (influenza virus) can help to evaluate the efficacy of antiviral drugs in inhibiting viral replication, enhance mucociliary clearance, and elevate the innate immune defence of airway epithelium.


Confocal image of 3D nasal and bronchial models

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Confocal image of infected 3D nasal and bronchial models. Decrease in ciliated cells and increase in goblet cells observed.

Experimental design for drug testing using viral-infected nasal/bronchial models

   Test Model   Nasal/Bronchial models
   Replicates   N=3 tissues per test condition
   Exposure Time   24-hour topical exposure to a predetermined 4-concentration rage of test chemical

   24-hour exposure to virus

   Assay Controls   Negative Control – Sterile DI H2O

   Positive Control – MOI 10

   Endpoints   MTT, TEER, Marker staining, real-time PCR
   Data Delivery   Cell viability, permeability analysis,cell type and gene analysis

 

Wound healing and Inflammation models

The wound healing process involves the repairing of skin homeostasis. It is a complicated process involving multiple interlinked phases such as haemostasis, inflammation, proliferation, and remodelling which generally takes around 4 weeks.

Obstacles in wound healing can occur from internal factors such as diabetes, coronary artery disease, aging, stress and immune system related diseases, as well as external factors such as bacterial infections, medication, smoking and nutrition. If not treated properly, these factors may lead to chronic non-healing wounds and resulting complications such as infection and tissue necrosis.

Invitrocue provides in vitro 3D skin models for the preliminary efficacy and safety evaluation for wound dressings, cosmetic products and skincare products.

 

In vitro 3D wound healing models

Our in vitro 3D skin models are composed of normal human epidermal keratinocytes (NHEK) and normal human dermal fibroblasts (NHDF)—cell types that mimic the wound bed. Interaction of keratinocytes and fibroblasts is important during the rebuilding of tissue integrity. Fibroblasts play an important role in wound healing by supporting the expansion and migration of keratinocytes.

Our 3D skin models are superior to the conventional monolayer model as they better recapitulate in vivo epidermis and skin barrier functions.

Advantages of our model:

  • More complex model than the 2D scratch wound assay
  • Cells are grown in a 3D collagen matrix that can provide structural support for the skin model
  • Allows for the study of cell migration toward the wound bed with the use of 2 major cell types (NHEK and NHDF)
  • Flexible model that can induce inflammation and mimic diabetic wound conditions by modifying the composition of the culture medium

Applications:

  • Wound dressing
  • Drug evaluation
  • Cosmetic products

The diagram above shows the method of wound preparation (gap creation) in our 3D co-culture of keratinocytes and fibroblasts. The wound healing and cell migration can be easily quantified through the measurement of gap closure over the time.

Reference: Jonkman, James E N et al. “An introduction to the wound healing assay using live-cell microscopy.” Cell adhesion & migration vol. 8,5 (2014): 440-51. doi:10.4161/cam.36224

   Test Model   3D co-culture model
   Replicates   N=3 tissues per test condition
   Exposure Time   Depends on the test materials
   Endpoints   Live/dead cell staining, cell migration evaluation
   Data Delivery   Cell viability, migration rate

 

For inquiries, please contact us.