Genes and Tissue Culture Technology

1. Potentials of Using 3D
In Vitro Models for Drug
Efficiency Testing
(in comparison to 2D)
By: TiffanyHo, ShannenSer, ArialChan, LimEeJing,OoJuNn

2. Basic Principles of 2D & 3D Cell Cultures
3D cultures
Growth condition Adhere and grow on a flat
surface
Grow on a matrix or in
suspension medium
Morphology Flat & stretched ; monolayer Form aggregates or
spheroids
Cell status Mostly proliferating stage Mixture of cells at different
stages
Limitation Does not adequately mimic
the in vivo microenvironment
Core cells receive less
oxygen, growth factors and
nutrients from medium ; in
quiescent or hypoxic state
Advantage Receive nutrients, growth
factors and oxygen equally
Mimic in vivo
microenvironment
Table 1 shows the comparison of principles of 2D & 3D cultures. (Edmondson 2014)

3. Potentials of 3D Cultures for Drug Testing

4. Applications
➔ Study of tumor development
- Cancer cells response to host immune modulatory effect.
➔ Evaluation of anticancer drug sensitivity
➔ Drugs discovery
➔ High throughput screening
➔ 3D cell-based biosensors
- Investigate cell’s response to drugs
- Detect biological signals transmitted by the cell
➔ Microfluidic-based device: Organs-on-chips
- Serve as disease model; mimic human organ functions.
- Small device with hollow channels lined by living cells cultured with nutrient liquids flowing
through the channels.
Figure 1: How biosensors work (Vaghasiya n.d.)

5. Cancer cells response to host immune modulatory effect
γ
γ

6. Drug Discovery
● Oncology drug development
- 2D cell cultures may not accurately mimic the 3D environment
- Fundamental differences in the microenvironment of 2D and 3D cell cultures influences cellular
behaviours
- Crucial difference : Dissimilarity in cell morphology
> 2D: Monolayer
> 3D: Aggregrates and Spheroids
● 3D have higher ability to show reliable data especially when it comes to drug testings
Figure 3: Cell
morphology of 2D and
3D

7. High Throughput Screening (HTS)
● HTS assays using monolayer (2D) cultures still reflect a highly artificial cellular environment
- Limiting the predictive value for the clinical efficacy of a compound
● Optimize preclinical selection of the most active molecules from a large pool of potential effectors
● 3D cell culture systems:
- Spheroids are emphasized due to their advantages and potential for rapid development as HTS
systems
- 3D double network Hydrogels are similar to natural tissues and their chemical tunability which
impart abilities for response

8. ❏ Mimics lungs physiology.
❏ Translucent design allows the viewing of inner workings of
human lungs.
❏ Contains tiny hollow channels lined with lung cells and
capillary cells separated by porous membrane.
❏ Vacuum pumps on either side of each channel expand and contract,
thus imitating the action of a real alveolar sac. (Whitwam 2012)
Figure 4: Lung-on-a-chip (Anthony 2012)
Figure 5: Lung-on-a-chip as model for pulmonary edema
(Whitewam 2012)
❏ Mimics inflammatory response triggered by microbial
pathogens.
e.g. WBCs migrate across capillary cells into the air space to
engulf bacteria.
❏ Used to model pulmonary edema by introducing IL-2 in
blood channel.

9. Biomaterials Technology
● Explores materials which are not passive and walled off by the body
● Actively participates in body’s effort to repair itself
● Biometric and bioactive materials are designed to mimic the body’s natural structures &
functions from macro- to micro- to nano-levels.
● Give rise to tissue and organ development
● Replacement of animal testing using combined models
Figure 6:
Schematic illustration of tissue
engineering based on 3D biomaterials
technology.
- Regeneration of defective and
injured tissue
Current Developments

10. 3D Printing Technology
● Biological construct in small range
(mm-cm), including several cell
types and biomaterials at the same
time
● Use 3D biomaterials printing and
with cell patterning
● Constructing 3D scaffolds with living
cells embedded in hydrogels
● Functional tissue is formed faster
compared to classical tissue
engineering methods
Figure 7: 3D printing technology for tissue engineering

11. Thank You!
Questions?

12. References
Antoni, D, Burckel, H, Josset, E & Noel, G, ‘Three-Dimensional Cell Culture : A breakthrough in Vivo’, International Journal of
Molecular Science, vol. 7, no. 1, viewed 28 April 2016, <http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4394490/>.
Dougherty, E 2010, Living, breathing human lung-on-a-chip: A potential drug-testing alternative, viewed 28 April 2016, <http://wyss.
harvard.edu/viewpressrelease/36/living-breathing-human-lungonachip-a-potential-drugtesting-alternative>.
Edmondson, R, Broglie, J, Adcock, A & Yang, L, ‘Three Dimensional Cell Culture Systems and Their Applications in Drug Discovery
and Cell-based Biosensors’, International Journal of Molecular Science, vol. 3, vol.1, viewed 28 April 2016, <http://www.ncbi.nlm.nih.
gov/pmc/articles/PMC4026212/#B5>.
Tolikas, M 2014, Wyss Institute's technology translation engine launches 'Organs-on-Chips' company, viewed 28 April 2016, <http:
//wyss.harvard.edu/viewpressrelease/161>.
Ou, K & Hosseinkhani, H, 2014, ‘Development of 3D in Vitro Technology for Medical Applications’, IJMS, vol. 15, no. 10, pp.17938-
17962.
Vaghasiya, K, Applications of Biosensors technology : Future trends development and new intervation in biotechnology, viewed 28
April 2016,<http://www.pharmatutor.org/articles/applications-of-biosensors-technology-future-trends-development-and-new-intervation-
in-biotechnology>.
Whitwam, R 2012, Lung-on-a-chip could change the way disease is treated, viewed 28 April 2016, <http://www.geek.com/chips/lung-
on-a-chip-could-change-the-way-disease-is-treated-1527521/>.