cell cultur

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