1. 3-D Cell Culture

2. Know these terms !! Extra Cellular Matrix ( ECM )
Extra cellular molecules secreted by cells for mechanical support, adhesion of cells, cell to cell interaction etc…
Basement membrane is an ECM

3. Scaffolding
A temporary structure made of wooden planks or bamboo made while construction of building

4. Scaffolding in Biology
It is used for Tissue regeneration and in-vitro drug testing.
Scaffolding of human ear

5. A 3-D, biodegradable, synthetic scaffold molded into precise shape of hip joint.
Known as Ghost heart
+ stem cell of recipient
Functional heart
Bioreactor

6. 2-D Cell Culture
Allows growth only in 2 dimensions in petridish ( mono layered )
Eg: Bacterial or yeast culture we do in our labs
Yeast transformation
Phage titration

7. Then why should we develop 3-D cell culture.
Advantages
Easier environmental control, cell observation and measurements
Cheaper to cultivate
Highly efficient in studying unicellular organisms
Then why should we develop 3-D cell culture.
In simple words, all our ( Human ) & animal cells are in 3-D environment. Culturing these cells in 2-D monolayer manner is a poor model for in vivo conditions and behaviors. ( Why ???!! )

8. Can tumor cells grown in 2-D mimic these in-vivo conditions ?
Different regions of a Tumor spheroid
Can tumor cells grown in 2-D mimic these in-vivo conditions ?

9. Let’s see an example !!! Human epithelial carcinoma cells
( A431.H9 cells )
5-fluorouracil ( 5 FU)
Grown in 2-D as well as 3-D conditions
Treated with 2 anti cancer drugs with diff. activity profiles
Tirapazamine ( TPZ )

10. Which result ( 2D or 3D ) is more reliable ???
Observation :A431.H9 cells are more resistant to
5-FU in 3D conditions than 2D
TPZ in 2D conditions than 3D
Which result ( 2D or 3D ) is more reliable ???

11. Why 2-D is a poor model for studying animals ?
Mono layered
ECM
Cell to cell interaction Absent
Cell & matrix interaction
Cells cannot attain their in vivo configurations as in tissues
Due to these reasons 2D cells gives inaccurate data regarding predicted response of cancer cells to chemotherapeutics.
 In 2011 alone, out of approximately 900 anti-cancer therapies in clinical trials or under Federal Drug Administration review, only 12 achieved approval ; resulting in the loss of hundreds of millions of dollars that were spent on pre-clinical and clinical trials. 

13. 3-D Cell culture models Scaffold based Non Scaffold based
Polymeric Hard Scaffolds
Biological scaffolds
Micropatterened surface
micro plates
Hanging drop micro plates
Spheroid micro plates with Ultra low attachment (ULA ) coating
Microfluidic 3D cell culture

14. Commonly we use Poly styrene , Polycaprolactone
Polymeric Hard Scaffolds
3-D culture made by growing cells on pre fabricated scaffolds made of polymers that mimic ECM.
Commonly we use Poly styrene , Polycaprolactone

15. Scaffolds created of components of biological origin.
Biological Scaffolds
Scaffolds created of components of biological origin.
Eg: fibronectin, collagen, laminin, gelatin
Collagen matrix
Silk Scaffold
Micro patterned surface micro plates
(A) round, (B) square, or (C) slit patterning within the plate well

16. What determines the size of spheroids ??
Hanging drop micro plates
Cells in absence of a substrate to attach with will self assemble to form spheroid structure
HDP top and bottom openings
What determines the size of spheroids ??

17. Spheroid micro plates with ULA coating
These plates can contain larger volume of medium than HDP
Microfluidic 3D cell culture
Accounts complexities such as continuous nutrient flow, waste removal etc…

18. Can you imagine some other method ???

19. Questions to Ponder…
What are some challenges / limitations of 3D cell culture?
Can traditional assays developed for 2D models be applied in 3D?
Does humans/ animals have higher dimension than 3D ?

20. Bibliography www.biotek.com https://www.ncbi.nlm.nih.gov/

21. Vishnu Narayanan S S BS-MS 2015