EQUIPMENTS USED IN CELL CULTURE (M PHARM SEM 1 (PH.COLOGY) UNIT 1 CMB

1. CELLULAR AND MOLECULAR BIOLOGY
19-01-2022 1
DEPARTMENT OF PHARMACOLOGY
Faculty of Pharmaceutical Sciences
PES University
HN Campus

2. TITLE: EQUIPMENTS USED IN ANIMAL CELL
CULTURE MEDIA
PES3PG21PL003
10-02-2024 2
Under the Guidance of:
Mr. Vimal john samuel
FACULTY OF PHARMACEUTICAL SCIENCES
PES University
PRIYASHREE K

3. Cellular and molecular biology
10-02-2024 3
CONTENTS
Introduction to Cell culture
History of cell culture
Equipments used in cell culture
Reference
Equipment's used in Animal cell culture

4. Equipment's used in Animal cell culture
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INTRODUCTION
 Animal cell culture is the removal of cell, tissue or organs from an animal and its
subsequent placement into an artificial medium conductive for growth.
 Animal cell culture are used in the production of therapeutic proteins, research
and diagnosis of diseases.
Cellular and molecular biology

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History of animal cell culture
 1885 – Wilhelm Roux first time maintained embryonic check cell in saline buffer.
 1903- Jolly first time showed that the cells can survive and divide in In vitro
condition.
 1907- Ross Harrison developed a nerve fiber from frog embryo tissue and culture
it in a blood clot.
 1912- Alexis Carrel evolved the cell culture by developing the first cell line.
Equipment's used in Animal cell culture
Cellular and molecular biology

6. Equipment's used in Animal cell culture
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 1940- Enders, Weller and Robbins grew the poliomyelitis virus.
 1955- Eagle developed a defined medium for cell growth.
 1964 – Little Field discovered the HAT medium for cell selection.
 1965- Ham discovered the first serum free media.
 1965- Harris and Watkins fused the first human and mouse cell by using a virus.
Cellular and molecular biology

7. Equipment's used in Animal cell culture
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EQUIPMENTS REQUIRED FOR ANIMAL CELL CULTURE:
 Laminar Air Flow Hood
 Co2 Incubator
 Centrifuge
 Refrigerator
 Cryo-storage container
 Liquid nitrogen container
 Haemocytometer
 Water bath

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 Flow cytometry
 Inverted Microscope
 Dissecting microscope
 Glass wares and plastic wares
Cellular and molecular biology

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BIOSAFETY CABINET
 The basic purpose of Biosafety Cabinet is to
provide protection to the culture from any
organism such as fungi, virus, bacteria under
aseptic conditions and also protect the
operator form the risk of infection.
 The purpose of this cabinet is to protect the
product by use of HEPA filters (High
Efficiency Particulate Air).
 The level of contamination depends on the
use of biosafety cabinet.

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1.BSC 1= Should have a biosafety cabinet, Efficiency of HEPA filters is 90%
2. BSC 2= Closed Biosafety cabinet, Efficiency of HEPA filters is 95%.
3. BSC 3= Closed Biosafety cabinet, High density, Radioactive sterilization,
Efficiency of HEPA filters is 99.5%

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CO2 INCUBATOR
 Incubator is a temporary chamber which provides suitable
temperature, density, moisture and pH for better growth
and development of animal cells.
 They maintain sterility of the chamber by using HEPA
filters.
 They maintain a constant temperature by using a silicon
gasket on the inner door.
 Humidity is created which prevents desiccation of the
media and maintain osmolarity.
 pH is also maintained in the media.

12. Equipment's used in Animal cell culture
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COOLING CENTRIFUGE
 Centrifuge is an instrument used to separate the
particles from a solution according to their size,
shape, density, viscosity of the medium and the
rotor speed by using the principle of centrifugal
force.
 Cooling centrifuge is used to prepare the cell for
cryo-preservation.

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REFRIGERATOR
 Cell culture laboratories needs a domestic
refrigerator (preferably one without an auto
defrost freezer).
 It stores the reagent, media and cells at 2-8
degree centigrade used for small laboratories.
 For large laboratories a cold room is restricted
for the storage of cells.

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:
CRYO-STORAGE CONTAINER
 Cell lines in continues culture are likely suffer
form genetic instability as their passage number
increases therefore it is essential to prepare
working stocks of the cells and preserve them in
cryogenic storage.

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LIQUID NITROGEN CONTAINER
Liquid nitrogen container provide a long term
storage with low liquid nitrogen evaporation.
There are two main types of liquid nitrogen storage
system :
1. vapour phase- this system minimizes the risk of
explosion with cryo-storage tubes.
2. liquid phase- This system usually have longer
static holding time.

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HAEMOCYTOMETER
It is an instrument used to estimate
the cell number and viability.
It has two parts:
1. Neubauer’s slide- it has counting
scales on each quadrant. The
depth of the scales is 0.1mm.
2. Coverslip – it is used to keep the
liquid sample in a perfect shape into
a flat layer of even thickness.

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WATER BATH
 A water bath is a device that maintains water at
a constant temperature.
 It is used for thawing of the cells directly taken
out from the freezing media or from liquid
nitrogen to prevent the cell form any
mechanical damage due to fluctuation of
temperature.
 The temperature of water bath is 5-100 degree
centigrade.

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FLOW CYTOMETER
o Flow cytometry is a laser based technique to
analyse the characteristics of the cells .
o This is mainly used for the analysis of the
expression of the cell surface and intracellular
molecules.
o It is used to characterize different cells from a
heterogenous mixture.

19. Equipment's used in Animal cell culture
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INVERTED MICROSCOPE
 The inverted microscope is designed with the
light source and the condenser lens above the
specimen.
 The objective and turret of the microscope is on
the bottom.
 The objective focus the light to produce a real
image.
 It is to observe any contamination in the cell
culture.

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Equipment's used in Animal cell culture
DISSECTING MICROSCOPE
 The dissecting microscope is also know as stereo-
microscope.
 It has a long working distance between 25 and
150mm.
 It has low magnification ability and one can
manipulate the specimen even performing a small
dissection under the microscope.
 Light specimens can also be observed under this
microscope.

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GLASSWARE's AND PLASTIC WARES
 Beaker
 Pipette
 Measuring cylinder
 Horizontal and vertical flask
1. T25
2. T75
3. T100
 Petri plate
 Microscopic slides
 Coverslips
 Cell culture plates.

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REFERENCE
 Culture of animal cell, manual basics techniques, R. Ian Freshney.
 Animal cell culture and technology, M. Butler.
 A text book of animal biotechnology, S.K Gautam, B. Singh, M S Chauhan.
 https://www.slideshare.net/SubhalaxmiSwain1/equipments-used-in-
animal-cell-culture

23. THANK YOU
Priyashree k
Department of pharmacology
10-02-2024 23