Alternative animal experimentation technique

reference

1. KARNATAKA COLLEGE OF PHARMACY DEPARTMENT OF PHARMACOLOGY PHARMACOLOGICAL AND TOXICOLOGICAL SCREENING METHOD PRESENTATION OF ALTERNATE ANIMAL EXPERIMENT     P REPARED BY : SUDARSHAN SINGH M pharmacy ( dept; pharmacology)

2. CONTENT Limitation of animal experimentation Alternate animal experiment Screening of alternate animal experiment

3. Limitation of animal experimentation Animal research and testing to protect and improve human health is not only unsafe But also expensive, time-consuming, and unreliable Animal models to study human diseases. Species differences in anatomy, organ structure and function

4. Examples penicillin is toxic to guinea pigs aspirin is poisonous to cats

5. Animal Alternatives: It is not possible to replace whole animal models with in vitro systems to evaluate drug effects on major organ systems

6. Alternative techniques: definition The term ‘alternative’ is used to refer to those techniques or methods that replace the use of laboratory animals altogether, reduce the numbers of animals required,or procedure or technique to minimize the level of stress endured by the animal.

7. Screening methods of alternate animal experiment 1) Full thickness skin model ( invitro method)*¹. 2) In sillico methods*². 3) cell line technique. 4) Patch clamp method

8. Full Thickness Skin Model: Robust full thickness skin model A test substance is applied to the Full Thickness Skin Model so that its effect on the skin tissue can be systematically evaluated. The substance, e.g. a cream formulation, is applied topically using a brush

9. done several times over a period of at least nine days. substance on the cell layers in the skin can be studied The standardized production of the model, in combination with its special properties, make it suitable for use as an in-vitro alternative to animal testing.

10. In- sillico methods Substances with similar chemical structures often have similar properties knowledge of the properties of a few representative substances is sufficient to be able to deduce the properties of a series of similar substances. certain properties of these representative substances can also be assumed to be properties of the other substances in the series. The required calculations are performed using specially developed computer programs.

11. Cell line technique: The term cell line refers to the propagation of culture after the first subculture. Once the primary culture is sub cultured, it becomes a cell line A cell line derived by selection or cloning is referred to as cell strain. Cell strain do not have infinite life, as they die after some divisions.

12. Types of cell lines : 1) Finite cell line. 2) Continuous cell line.

13. (1) FINITE CELL LINE: The cell line with limited culture life spans are referred to as Finite cell line. The cell normally divide 20-100 times before extinction. The actual number of doublings depends on the species, cell lineage differences, culture conditions etc.

14. (2) CONTINUOUS CELL LINE: The continuous cell lines are transformed, immortal and tumorigenic . A few cells in culture may acquire different morphology and get altered. Such cells are capable of growing faster resulting in an independent culture. The progeny derived from these altered cells have unlimited life.

15. APPLICATIONS OF CELL LINE : Screening of anti cancer drugs Cell based bioassays In vitro screening of several drugs Production of anti viral vacci Gene therapy Recombinant DNA therapy Biotechnology

16. PATCH CLAMP TECHNIQUE Patch clamp technique is a technique in electrophysiology that allows the study of individual ion channels in cells. The technique is used to study excitable cells such as neurons, muscle fibers and the beta cells of the pancreas

17. Types of patch

18. Patch cell technique in kidney cell

19. Procedure The patch clamp technique can be applied to cultured kidney cells freshly isolated kidney cells or to cells of isolated perfused kidney tubules Segments of late superficial proximal tubules of rabbit kidney are dissected and perfused from one end with a perfusion system

20. Evaluation In isolated perfused renal tubules, concentration response curves of drugs which inhibit ion channels can be obtained with the patch clamp technique

21. Applications of Patch Clamp Technique: For the evaluation of antiarrhythmics agents. In kidney cells. Used for isolated ventricular myocytes from Guinea pigs to study a cardio selective inhibition of the ATP sensitive potassium channel

22. Thank you