This document discusses aseptic techniques and sterile handling in animal cell culture. It explains that all cell culture procedures must be done aseptically under controlled conditions, such as a laminar flow hood. Basic aseptic techniques include cleaning the work area with ethanol and sterilizing all equipment. Materials used in cell culture must be carefully sterilized, with heat-stable components autoclaved and heat-labile materials filtered. Autoclaving is the preferred sterilization method as it is cheaper and more effective than filtration. Safety precautions for cell culture include keeping cultures antibiotic-free and never sharing media between cell lines.

1. ASEPTIC TECHNIQUES AND
STERILE HANDLING IN
ANIMAL BIOTECHNOLOGY
PRESENTED BY:
AKSHDEEP SHARMA
REG NO: 12BTB003
B TECH BIOTECH
6TH SEM

2. CELL CULTURE IS THE COMPLEX
PROCESS BY WHICH CELLS ARE
GROWN UNDER CONTROLLED
CONDITIONS. IN PRACTICE, THE
TERM "CELL CULTURE" HAS
COME TO REFER TO THE
CULTURING OF CELLS DERIVED
FROM MULTICELLULAR
EUKARYOTES, ESPECIALLY
ANIMAL CELLS .
Animal Cell/Tissue Culture:

3. Aseptic techniques :
 In culturing animal cells, it is essential
that all procedures are carried out
using aseptic or sterile techniques.
Laminar flow facilities or sterile rooms
provide a suitable environment, but
even then aseptic techniques should
be employed.
 Basic aseptic technique is to ensure
that the work area is clear, swabbed
down regularly with 70% ethanol and
that all the equipment used has been
sterilized.

4. A: Performing all procedures in a
laminar flow hood.

6. B: Using flames to fix
microorganisms on container necks

7. C: Holding a bottle
cap with the little finger.

8. D: Avoid touching tops of open
vessels while transferring
their content.

10. Aseptic techniques :
 Avoiding placing caps & pipettes
down on the bench; practice holding
bottle tops with the little finger.
 Work either left to right or vice versa,
so that all material goes to one side.
 Clean up spills immediately & always
leave the work place neat & tidy.

11. Aseptic techniques :
 Switch on UV light at
least 20 minutes before
and after use.
 Avoid placing pipettes
upside down.
 Use gloves, masks and
lab coats while
working.

12. Sterilization :
 Sterilization (or sterilisation) is a term
referring to any process that
eliminates (removes) or kills all forms
of life, including transmissible agents
(such as fungi, bacteria, viruses, spore
forms, etc.) present on a surface,
contained in a fluid, in medication, or
in a compound such as biological
culture media.

13. Sterilization Technique:
 The various media constituents and
other reagents used in cell cultures
must be carefully sterilized either by
autoclaving or by filtration.
 Heat stable constituents like water,
salts , supplements like peptone or
tryptose, etc. are autoclaved at 121°C
for 20 minutes.

14. Sterilization Technique:
 Heat labile constituents like serum,
trypsin, proteins, growth factors etc.
must be sterilized by filtration through
a 0.2 micron porosity membrane filter.

15. Autoclaving is preferred to
filtration:
 Autoclaving is preferred to filtration
since it is cheaper.
 Autoclaving needs less labour.
 Autoclaving is uniformly effective.
 Most of the media, now available
commercially are usually presterlized.

18. Safety aspect in cell culture:
 Possibly keep cultures free of
antibiotics in order to be able to
recognize the contamination.
 Never use the same media bottle for
different cell lines.
 Necks of glass bottles prefer heat at
least for 60 secs at a temperature of
200°C.

19. Safety aspect in cell culture:
 Switch on the laminar flow cabinet 20
minutes prior to start working.
 Cell cultures which are frequently used
should be subcultered & stored as
duplicate strains.