Cell Culture

Primary and Continuous Cell Line Culture

• Primary culture: Cells freshly isolated from tissues.

• Continuous cell lines: Immortalized cells that can be cultured indefinitely.
   

Cell Viability Assays (Trypan Blue, Resazurin)

• Trypan blue exclusion: Stains dead cells to estimate viability.

• Resazurin assay: Measures metabolic activity of living cells.
   

Toxicity Testing of Drugs/Compounds (MTT Assay)

Determines cytotoxic effects of compounds by measuring mitochondrial activity in cells.
 

Apoptosis Detection (Annexin V/PI, Caspase Assays)

• Annexin V/PI staining: Differentiates early apoptotic, late apoptotic, and necrotic cells.

• Caspase assays: Detects activation of caspase enzymes involved in programmed cell death.
   

Antibody Production (Monoclonal/Polyclonal)

• Monoclonal antibodies: Produced from a single clone, highly specific.

• Polyclonal antibodies: Mixture from multiple clones, broader reactivity.
   

Transfection (Lipofection, Electroporation)

Introduction of DNA, RNA, or proteins into cells for gene expression or silencing studies.
 

3D Cell Culture and Spheroid Models

Culturing cells in three-dimensional systems to better mimic natural tissue environments.
 

Immunocytochemistry and Staining

Detection of proteins or biomarkers within cells using labeled antibodies and microscopy.
 

Co-Culture Systems (Tumor-Stroma, Immune-Tumor)

Culturing two or more cell types together to study interactions, such as cancer-immune or cancer-stroma relationships.
 

Cell Migration and Invasion Assays

• Migration assays (e.g., wound healing/scratch assay): Measures movement of cells.

• Invasion assays: Test cancer cell ability to invade through extracellular matrix.
   

Stem Cell Differentiation Assays

Inducing stem cells to differentiate into specific lineages (neurons, adipocytes, osteocytes, etc.).
 

Cytokine Profiling (Luminex, ELISA)

Quantifies cytokines released by cells, useful in immunology and inflammation studies.
 

Autophagy and Necrosis Detection

• Autophagy assays: Detect cellular self-degradation processes.

• Necrosis assays: Identify uncontrolled cell death caused by damage or stress.
   

Karyotyping and Chromosomal Aberration Studies

Microscopic examination of chromosomes to detect genetic abnormalities.
 

Cell Cycle Analysis

Flow cytometry-based measurement of DNA content to determine distribution of cells across G0/G1, S, and G2/M phases.
 

Mycoplasma Contamination Testing

Detection of mycoplasma (common contaminant in cultures) using PCR or fluorescence staining.
 

Clonogenic Assay

Tests the ability of a single cell to grow into a colony, used for radiation/drug sensitivity studies.

AlamarBlue Assay

Fluorescent assay to measure cell viability and metabolic activity, similar to resazurin