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Alternative animal experimentation technique
1. KARNATAKA COLLEGE OF PHARMACY
DEPARTMENT OF PHARMACOLOGY
PHARMACOLOGICAL AND TOXICOLOGICAL SCREENING METHOD
PRESENTATION OF
ALTERNATE ANIMAL EXPERIMENT
PREPARED BY : SUDARSHAN SINGH
M pharmacy ( dept; pharmacology)
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
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.
10. 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.
11. 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.
12.
13. 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.
18. (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.
19. (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.
20.
21. 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
22. 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
26. 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
27. Evaluation
In isolated perfused renal tubules, concentration
response curves of drugs which inhibit ion channels
can be obtained with the patch clamp technique
28. 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