PRESENTATION BY :

M.THILAKAR,LS1154,THIRD YEAR,M.SC . LIFE SCIENCES,BDU.

Cell..??

• Basic unit of Life.• Prokaryotic cells• Eukaryotic cells

• Functions of cells.?• Organs.?• Organism.?

CELL CULTURE

• Culture of living cells in the in vitro condition with suitable growth media.

• Should need • Optimum Temperature,• pH,• Proper media,• Incubator

• Out of Microbial contamination , etc.

Revision..• Types of Culture.?

• Cell culture,• Tissue culture,• Organ culture

• What is Callus.?• Dedifferentiation..?

• Differentiation..?• Redifferentiation.?

• Uni potent.?• Multi potent.?• Primary Vs. Secondary culture (Cell lines)..?• Sensence.?

Tumor

• Uncontrolled growth of normal cells.• Why.?• Gentical,• Environmental,• Other Body regulatory factors.

Tumor cell

culture

Tumor Cell Culture• Primary tumor cell culture.?

• Sampling of tumor cells.• Initial establishment.• Disaggregation.

• Cancer cell lines.?• Characterization of Tumor cells.• Common Problems with Tumor cell culture.

Primary tumor cell culture.?

• Sampling of tumor cells.• Cell banks.

• World federation for Culture – Having 469 centers in world level.

• Isolation of Tumors from Animals.• Initial Establishment.

• Transfer of Fragment to hood.• Viable cells. (Remove Necrotic cells)• PBS wash (To remove Blood)

Disaggregation

• Mechanical method & • Enzymatical method.

Mechanical

• Scalpels,• Pulverize,• Avoid cluster of cells,• Centrifuge,• Inoculate.

Enzymatical

• Trypsin,• Collagenase,• Elastase,• Dispase,• Hyaluronidase,• Papin.

CHARACTERIZATION OF TUMOR CELLS

• While establishing new cell lines, it is required.

• Origin of cell lines.• 3 Methods :

• Access uniqueness• Specific origin• General features

1.Access uniqueness

• Using Molecular biological experiments.• Based on DNA profiles.

• DNA finger printing,• DNA profiling,

2. Species of Origin

• Iso enzyme analysis• Cytogenetic studies,• Antibody detection methods (IFA),• PCR studies.

3.General features

• Morphological studies,• Cultural characters, • Cell cycle characters,• Cell – Cell communication.

My References

1) Methods in Molecular medicine – Cancer Cell culture – Methods & Protocols by Simon P.Langdon, Humana press.

2) Primary cell culture guide – Tips & Techniques – ATCC.