National Centre for Cell Science

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National Centre for Cell Science
Service
Research
Teaching/ Training
Design of the Centre
Research Laboratories
Infrastructure
Establishment
Cell Biology
Washing / Packing
Protein Chemistry
Media
Molecular Biology
Repository
Parasitology
FACS/ Photography
Immunology
Computer/ Animal House
Liquid N2 Plant
Tissue Banking STANDARDISATION OF PROTOCOLS TO
CRYOPRESERVE GROW IN VITRO TISSUES
IMPORTANT IN TRANSPLANTATION BIOLOGY like
Cornea, Skin, Bone Marrow, Islets etc.
TRANSFER OF THE PROTOCOLS TO HOSPITALS
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Sources of Haematopoietic stem cells
MOUSE HUMAN
ADULT B.M. CORD BLOOD FETAL LIVER
Isolation Of Mononuclear Cells
Large Volume
Small Volume
FICOL/ HYPAQUE
HES
MNC FREEZING IN CRYO VIALS /CRYO BAGS
PROGRAMMABLE FREEZER
Program Section Function Temperature 1 Wait 4oC
2 2 1oC/ min -40oC 3 10oC/min -90oC 4 END
LOCATION IN LIQUID NITROGEN-196OC
THAWING IN WATER BATH AT 37oC
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PRE AND POST CRYOPRESERVATION PARAMETERS
1. NUCLEATED CELL RECOVERY
2. DYE EXCLUSION VIABILITY
3. CFU ASSAYS
Differential Colony Count
BFU (E)
GM
GEMM
TOTAL
HPPCFC
4. MORPHOLOGICAL IDENTIFICATION OF CELL TYPES
5. PHENOTYPIC ANALYSIS ON FLOW CYTOMETER
6. LTBMC
CFU (S)
7. IN VIVO (Murine)
ENGRAFTMENT
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RESEARCH
INNOVATION OF TECHNOLOGY OF FREEZING
MEMBRANE STABILIZERS
1. ADDITIVES TO CRYO - MEDIUM
BIOANTIOXIDANTS
AFP - AFGP
2. REDUCTION IN DMSO
- 196oC
3. STORAGE TEMPERATURES
- 80oC
4. GROWTH FACTOR RECEPTOR STATUS
5. ENGRAFTMENT/ HOMING POTENTI AL
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ACKNOWLEDGEMENTS
Dr. Anand Rao
Dr. V. Kochupillai
Dr. R. L. Marathe
Dr. Dilip Ghaisas
Sassoon Hospital
BTD A . F. M. C
Dr. V. P. Kale