Freshly removed cancer tissues were collected, a portion of which were cryopreserved and the rest processed in the laboratory. The laboratory was able to establish primary cultures of the following tissues: vaginal mucosa, cervix, uterus, ovary, breast mastoid, urinary bladder, kidney, thyroid, pineal body and brain. Vigorously growing cultures were passage several times and cryopreserved. Preliminary characterization of these cultures was done through inverted light microscope to observed its morphological characterization. Immuno cytochemical analysis was also done using monoclonal antibodies against intermidiate filaments like vimentin, cytokeratin 4, epidermal membrane antigen (EMA) and glial fibrillary acidic protein (GFAP). Paraffin-embedded tissues were also characterized immunohistochemically using the same antibodies. Cells were also tested with thymidine kinase antibody (TK-1) which is a marker of proliferation. Additional characterization will be done using mycoplasma test and karyotyping of the cell cultures. The project has recognized five primary cultures that are considered potential cell lines pending studies on their survival capabilities. These are uterus, breast, pineal gland, astrocytoma and meningioma.
GENERAL: 1. To establish cell lines from primary cultures of various human cancer tissues for the first project year;2.To develop a cell bank of cancer cells lines for biomedical researchers.
SPECIFIC: 1. To collect human biopsy materials/surgical explants from clinically diagnosed cancer patients;2. To establish primary cultures of specific cancer tissues from surgical explant, e.g. cancerous epithelial tissues;3. To develop cancer cell lines; 4. To characterize the cultivated cell lines in terms of a. growth patterns and morphology b. surface marker c. isoenzyme profile d. DNA fingerprinting e. cloning efficiency; 5. To formulate strategies for the distribution and widespread utilization of these materials.
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