Development of cancer therapeutics is often carried out in 2D cultures prior to testing on animal model. In comparison to 2D cultures, discuss the potential of using 3D in vitro models for drug efficiency testing.

1. Genes & Tissue Culture
Presentation
(Group 4)
By:
Asiah Salleh
Chua Qin Ling
Ng Chu Xin
Lim Su Shen

2. Development of cancer therapeutics is often
carried out in 2D cultures prior to testing on
animal model. In comparison to 2D cultures,
discuss the potential of using 3D in vitro
models for drug efficiency testing

3. General Introduction of 3D & 2D culture

4. Video Introduction

5. Comparison between 2D and 3D culture

6. 2D/3D Culture in drug efficacy testing
➔ Studies have found that cells cultured in 3D models are more resistant to
anticancer drug than in 2D cultures
- geno- or phenotypical change induced by 3D spheroid formation
- difference in gene expression
- different cell stage
➔ 2D culture is non-predictive for in vivo test
- Drug candidates fail in clinical trials due to adverse events, low efficacy etc.
➔ 3D culture emerged as a more reliable model to test for cancer cell
viability in response to drug treatment
- Photodynamic therapy
(Zang et. al 2012)
(Edmondson et. al 2014)
(Chen et. al 2015)

7. Differences in efficacy of PDT between 2D monolayer cell cultures and 3D
spheres
● Perform Photodynamic Therapy
(PDT) to compare conventional
2D culture and 3D culture
● LIVE/DEAD staining to check for
cell viability
● After 10 minutes, about 50% of
the cells in 2D culture are viable,
but most of the cells are still
viable in 3D spheroid
● After 1 hour, all cells in 2D
culture were killed, but many cells
in the sphere are still viable
(Chen etl al 2015)

8. Advantages of 3D cell culture systems for drug
discovery.
★ Matrices contain ECM components lead to better cell-cell contact,
communication and signalling pathway activation.
★ Restored cell functional and morphological differentiation.
★ Culture condition can be modified to include factors/proteins found
in particular tumour microenvironment.
★ The gene and protein expression levels of cells as well as the
cellular behaviours are similar to the in vivo levels.
★ Provide in vitro models for including different types of cells to build
multicellular systems.
★ Bridges the gap between in vitro and in vivo drug screening,
(Edmondson et. al, 2014)
Figure : Advantages of microenvironment for drug
discovery programmes. (Lovitt et. al, 2014)

9. Disadvantages of 3D cell culture systems for drug
delivery
❖ variability in biologically derived matrices leads to non-
reproducible experimental results
❖ high cost needed for large scale studies and high throughput
assay
❖ High variability result ( difference ECM components in
matrices)
❖ Low throughput in certain technique, due to the necessity of
manual media changes (Katt et. al 2016)
(LabAutopedia, 2016)
Figure : Hanging Drop Plate Assembly.

10. Application of 3D cell culture
· study basic mechanisms of organ development
· to develop artificial organs for replacement or support of natural organs
· to optimize bioproduction, or to obtain realistic pharmacotoxicological test systems.
Drug discovery:
Undergo high throughput screening to generate hits.
Useful in cancer drug research
organotypic tissue or slice cultures for drug testing:
artificial liver (hepatocyte spheroids)
hormone-producing tissues
brain cell cultures
heart cells
extracellular matrix and skin (by using fibroblasts and/or keratinocytes)
Artificial skin

11. Embryoid Bodies
Embryoid bodies are derived from embryonic stem cell lines
retained capacity of lineage commitment (i.e., of generating cells of the hematopoietic,
endothelial, muscle, and neuronal lineages)

12. References
Chen, YC, Lou, X, Zhang, Z, Ingram, P & Yoon, E 2015, ‘High Throughput Cancer Cell Sphere Formation for Characterizing the Efficacy of
Photodynamic Therapy in 3D cultures’, Scientific Reports, viewed 29 April 2016,
<http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4495468/>
Edmondson, R, Broglie, JJ, Adcock, AF &Yang LJ 2014, ‘Three-Dimensional Cell Culture Systems and Their Applications in Drug Discovery and
Cell-Based Biosensors’, Assay and Drug Development Technologies, vol 12, no. 4, pp. 207-218.
Katt, ME, Placone, AL, Wong AD, Xu, ZS, Searson, PC 2016, ' In Vitro Tumour Models: Advantages, Disadvantages, Variables and Selecting the
Right Platform', Frontiers in Bioengineering and Biotechnology, vol. 4, no. 12.
Kunz-Schughart, L 2004, ‘The Use of 3-D Cultures for High-Throughput Screening: The Multicellular Spheroid Model’, Journal of Biomolecular
Screening, vol 9, no.4, pp.273-285.
LabAutopedia 2016, Automated Cell Dispensing and Image-based Spheroid Formation Tracking, viewed 26 April 2016,
<http://www.labautopedia.org/mw/Automated_Cell_Dispensing_and_Image-Based_Spheroid_Formation_Tracking>
Lovitt, CJ, Shelper, TB & Avery, VM 2014, ‘Advanced Cell Culture Techniques for Cancer Drug Discovery’, Discovery Biology, vol 3, no. 2, pp. 345-
367.
Mueller-Klieser, W 1997, ‘Three-dimensional cell cultures: from molecular mechanisms to clinical applications’, American Journal of Physiology -
Cell Physiology, vol 273, no.4, pp.C1109-C1123.
Zhang,R, Li, D, Tang, IC, Wang, J & Yang, ST 2012, ‘Cell-Based Assays in High-Throughput Screening for Drug Discovery’, International Journal of
Biotechnology for Wellness Industries, vol. 1, pp. 31-51.

13. THANK YOU!