Cell lines introduction ,and invitro bioassays in Diabetes and Arthritis.

1. Presented By:
Heena Parveen,
Dept. of Pharmacology.
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2. CONTENTS
 Cell lines
• Handling of cell lines
• Maintenance of cell lines
• Storage of cell lines
 Invitro cell based assays for diabetes
 Invitro cell based assays for arthritis
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3. CELL LINES
 The culture produced by the cell or tissue
taken from an organism is termed as primary
culture.
 Cell line implies to the sequence of culture
obtained from the sub cultivation of the
primary culture.
 A cell line is a product of single layer cells that
are used for biological research.
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4. PRIMARY CULTURE
 Directly from cells or tissues.
 Collagenase or trypsin – breakdown
extracellular matrix.
 Media is supplemented.
 Incubation and growth.
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5. BOVINE PULMONARY ENDOTHELIAL
CELLS
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7. CULTURE PLATES
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8. MULTICHANNEL PIPETTES
DISPOSABLE PIPETTES
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9. HANDLING OF CELL LINES
 Cell lines may be contaminated with
microbes.
 They should be handled in accordance with
OSHA (Occupational Safety And Health
Administration).
 Personal protective equipment outer wear
like gloves, gowns, face masks etc. should be
used.
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11. HANDLING OF CELL CULTURE PLATES AND
BOTTLES
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12. MAINTENANCE OF CELL LINES
 Cell lines are grown as monolayer in culture
vessels.
 They are either maintained under continuous
illumination or in the dark.
 The standard growth conditions for all cell lines
are
 24 °C - 37o
C and 40 % rel. humidity
 Two independent sets of the whole collection are
maintained independently.
 Maintaining of cell lines is done by passaging.
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13. PASSAGING OF CELL LINES
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14. PASSAGING OF CELL LINES
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16. CO2 INCUBATOR
Maintains temperature, humidity,
 CO2, O2 contents.
Adjustable heaters – maintain temperature.
Incubator shelving is
 perforated – circulation
 of air.
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17. INVERTED PHASE MICROSCOPE
 Views objects from an inverted position.
 Views live specimens.
 Views living cells at the bottom of a large
container than on a glass slide.
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18. STORAGE OF CELL LINES
Cryopreservation:
 Preservation at low temperatures
 Generally solid CO2 and
liquid Nitrogen are used.
Cryoprotectives are also used.
Most commonly used cryoprotectives
are DMSO and glycerol.
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19. LIQUID NITROGEN TANKS
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21. HUMAN STEM CELL LINES
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23. 86
Rb+
EFFLUX MEASUREMENTS
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25. ACTIVITY ON ISOLATED MUSCLE
CELL
Principle:
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26. Procedure:
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29. FORMATION OF LEUKOTRIENE B4 IN
HUMAN WHITE BLOOD CELLS
INVITRO
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31. COX-1 AND COX-2 INHIBITION
Whole cell assays with osteosarcoma
cells and U937 cells
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33. REFERENCES
Text book of Drug Discovery and Evaluation
Pharmacological Assays, Second Edition by
H. Gerhard Vogel (Ed.) Page no. 738, 740, 1034,
1040.
 Text book of Pharmaceutical Biotechnology by S.P.
Vyas and V.K. Dixit Page no. 277, 278.
www.cabri.org/guidelines/plant/700.html
 http://www.protocol-online.org/cgi-bin/prot/view
 http://www.biontex.com/con_4_6_4/cms/upload/pdf/M
irmira_MP_e.pdf
http://catalog2.corning.com/Lifesciences/media/pdf
/Cell_freezing_protocol.pdf
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