This document discusses different cell culture models, including 2D and 3D cell cultures. 2D cell cultures utilize a flat monolayer that does not accurately mimic the in vivo environment. 3D cell cultures, including scaffold-based and scaffold-free cultures, better simulate the complex 3D environment and cell-cell interactions seen in vivo. Growing cells in 3D alters their proliferation, morphology, drug response, gene expression, and behavior in ways that more closely resemble the in vivo tumor microenvironment compared to traditional 2D cell cultures. 3D cell cultures are useful tools for applications like cancer research, drug development, and modeling of complex tissue environments.

1. Theoretical Models
Cell Culture))
By:A.Haghbin

2. Model:The set is the experiment or effect on which to measure
Invivo models
Animal models
Invitro models:
Cell culture
- 2D Cell culture
- 3D Cell culture

3. Invivo models
• Animal models
- living incubators in which human tumors have been grown
-extensive use for screening
-currently lack a qualifiedmode for many forms of deadly cancer

4. Why is cell culture used for?
• Model systems
• Toxicity testing
• Cancer research
• Virology
• Genetic Engineering

5. Types of cells
• Primary cells
• Semi-continuous cells
• Continuous cells

6. Types of culture
• 2D Cell culture
- Monolayer
• 3D Cell culture
-Scaffold-base
- Scaffold-free : spheroid

7. 2D Cell Culture
• flat monolayers submerged in media
• it’s simplicity
• this model can’t accurately depict and simulate the rich environment and
complex processes observed in vivo:cell signaling, chemistry or geometry
A SEM 2D cell culture image

8. 3D Cell Culture
Scaffold-base
• provide cell growth support
• present biocompatibility properties
• Example:
Hydrogels Scaffolds
bioglass or bioceramic Scaffolds
Non gel Polymer scaffold

9. 3D Cell Culture
Scaffold-free : spheroid
• bridge the gap between in vitro experiments used for discovery
and screening and in vivo experiments
• perfectly mimic in vivo cells behaviors and organization
• MCTS :Multicellular tumor spheroids

10. Scaffold-free 3D cells cultures techniques
• forced-floating method
• agitation based method
• hanging drop method

11. 2D vs 3D Cell Culture
• Physiologic cell-to-cell contact dominates
•Cells interact with extracellular matrix
(ECM)
• Diffusion gradient of drugs, oxygen,
nutrients, and waste
• Co-culture of multiple cell mimics
microenvironment
• Shows resistance to anticancer drug as in
vivo tumor
• Cell-to-cell contact only on edges
• Cell mostly in contact with plastic
• Cells contact extracellular matrix mostly on one
surface
• No gradients present
• Co-culture unable to establish
amicroenvironment
• Anticancer drug resistance is not seen

12. Growing Cells in 3D alters proliferation
and cell morphology
• changes in proliferation rate
ECM-dependent:changes in intracellular signaling and stromal cell
influence
• changes in cell shape that ultimately modify cellular function
apical-basal polarity

13. Growing cells in 3D reveals a more realistic
drug response
• increase resistance to chemotherapy
• recapitulate several mechanisms of drug resistance

14. Growing cells in 3D captures phenotypic
heterogeneity
• Heterogeneity
- morphological and functional changes
• Two theories exist to explain these differences
-clonal selection theory
-cancer stem cells (CSCs)

15. Growing cells in 3D changes gene expression
and cell behavior
• gene expression patterns
- change intracellular signal transduction
-changes in cell contact directly

16. • genetic changes has impacts on cell behaviors such as cell
migration and differentiation
• interrogated cell migration
- Boyden chamber systems
- monolayer scratch assays

17. Growing cells in 3D mimics the tumor
microenvironment
• cancer is a disease of not only the tumor cell but also the
surrounding microenvironment
• cancer-associated fibroblasts (CAFs)

18. 3D Cell Culture Application
• Microfluidics 3D Cell Culture :Organs-on-Chips

19. Draw curve to increase the volume of sphrocheids
• Fractionation in radiotherapy

20. Reoxygenation
Hypoxiccellratio
Tumor growth process

21. Thanks for your attention