ZO425 : Cell and Tissue culture

أ.1

The student's knowledge of the various techniques used in cell and tissue culture

أ.2

Knowledge of the equipment and chemicals used in the cultivation of cells and tissues

أ.3

Know the factors that affect the quality of cells and tissues

evaluation number

Evaluation methods

assessing weight.

percentage

comments

First evaluation

First written test

2 hours

25

25%

seventh

Second evaluation

Second written test

2 hours

25

25%

The tenth

Final evaluation

Written and practical test

2

50

50%

According to the schedule

المجموع

100 درجة

100%

References address

publisher

Release

Author

located

Course notes

Cambridge Press

Neoplastic and normal cells in culture 1991

Vasiliev J.M. and Gelfand I.M

Libraries

Help books

ù Academic Press. NY. London.

The transformed cell 1991

Cameron I.L. and Pool. T.B

Scientific journals

ù 6th Edition. WIELY-LISS.

Culture of Animal Cells: A manual of basic technique 2010

Freshney, R.I.

Scientific journals

Humana Press

Protocols for neural cell culture 2010

Doering, L.C

Other

International information network

scientific topic

number of hours

lecture

independent study

week

scientific topic

1.

1. Introduction to human and animal cell and tissue cultures:

· What is cell culture?

· Historical background.

· Safety aspects of cell and tissue culture:

o Risk assessment.

o Biohazards.

o Genetically modified organisms.

o Disinfection.

o Waste disposal.

o Safety equipment.

o Personal Protective Equipment (PPE).

o Safe laboratory practices.

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2

3

7

2

2. Importance of and progress in animal cell culture technology:

· Importance of cell and tissue cultures:

· Applications of cell culture.

· Equipment for the cell culture laboratory:

o Laboratory Design.

o Microbiological Safety Cabinets.

o Centrifuges.

o Incubators.

o Work Surfaces and Flooring.

o Plasticware and Consumables.

o Care and Maintenance of Laboratory Areas.

12

2

3

7

3

3. Culture media:

· Definition.

· Types:

o Natural media.

o Synthetic media.

4. The cell environment:

· Media:

o Basic constituents of media.

o Inorganic salts.

o Buffering systems.

o Carbohydrates.

o Amino acids.

o Vitamins.

o Proteins and peptides.

o Fatty acids and lipids.

o Trace elements.

o Preparation of media.

o Serum.

o Guidelines for serum use.

o Origin of serum.

· pH .

· CO2.

o Temperature

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2

3

7

4.

5. Classification of tissue cultures based on the origin of the cells:

· Primary culture (directly from animal or plant tissue):

o Monolayer.

o Suspension:

Hematopoietic cells.

Established (transformed) cell lines

Cells from malignant tumors.

· Three dimensional cellular structure:

o Histoculture.

o Organotype culture.

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2

3

7

5.

6. Other cell culture types:

· In vitro culture:

o Primary culture.

o Secondary culture.

o Culture from established cell lines.

· In vivo culture:

o From donor (dead body):

pancreas.

liver.

o Stem cells (embryonic, mature stem cells).

Cloning (in vitro transformation of animal cells).

12

2

3

7

6،7.

7. Chemical defined of specific growth factors:

· Growth controlled by component of the humoral medium:

o growth factors.

o nutrients.

o ions.

· Growth factors and other medium components:

o the effect of whole serum (examples).

o the effect of purified growth factors.

· Epidermal growth factors (EGF):

o human fibroblast cells.

o bovine endothelial cells.

o human endothelial cells.

· Fibroblast growth factors (FGF).

· Other growth factors present in serum:

o glucocorticoids.

o dexamethanose.

o insulin.

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4

6

14

8.

8. Anchorage dependence:

· Colony forming efficiency correlation with tumorigenicity cells.

· Anchorage dependence during circulatory metastasis.

· Possible mechanisms of anchorage dependence

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2

3

7

9.

· Density dependent inhibition (DDI):

o calculation of labelling indices.

o wounding of the culture.

o comparison of proliferation cells.

· Alterations of density dependent inhibition (in transformed cell line cultures).

· Possible mechanisms of DDI.

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2

3

7

10

9. Alteration of morphogenetic reactions:

· Altered spendopodia attachment reaction of transformed cells.

· Alteration of fibronectin and other matrix.

· Alteration of diverse surface proteins.

· Alteration of lipids.

· Secretion of proteases.

· Phenotypic transformations.

10. Interactions of cells with territories:

· Cell territories in vitro.

· The spread of normal cells on new territories

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7

11،12

11. Cell behavior:

· Modes of cellular migration (examples).

o long range migration (chemotaxis, hepatotaxis, galvanotaxis, contact guidance).

o short range migration:

contact inhibition of movements.

thermodynamic model of cell interaction (sorting out, random, separation).

· Role of cell surface:

o cell membrane.

o molecules under the cell membrane:

microfilaments, microtubules.

o molecules above the cell membrane:

cell adhesion molecules (CAM):

o general, restricted, saccharides.

cell junction molecules.

substrate adhesion molecules (SAMs):

o extracellular matrix component (collagen, proteoglycans, glycoproteins).

24

4

6

14

13

12. Sourcing of cell lines.

13. Authentication of cell lines.

14. Cryopreservation and storage of cell lines:

· Cryopreservation.

· Ultra-low temperature storage.

· Inventory control.

· Safety considerations.

12

2

3

7

14

15. Insect cell culture: an overview.

· Morphology of Sf21 cells.

· Morphology of Sf9 cells.

12

2

3

7