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Mammalian Cell Culture

Mammalian Cell Culture. Majeed Arsheed Sabbah , phD Biotechnology research center, Alnahrain university. What is cell culture. Current work in Iraq. Cells isolation. Ficoll. Immunomagnetic separation. Cell viability. Trypan Blue exclusion.

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Mammalian Cell Culture

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  1. Mammalian Cell Culture MajeedArsheedSabbah, phD Biotechnology research center, Alnahrain university

  2. What is cell culture

  3. Current work in Iraq

  4. Cells isolation Ficoll Immunomagnetic separation

  5. Cell viability Trypan Blue exclusion live cells exclude the dye as the membrane is impermeable, but dead cells uptake the dye due to the lack of an intact cell membrane. Viability is then determinedby manually counting the stained sample in a hemacytometer, a time consuming and labor intensive process. More importantly, trypan blue stained samples have to be analyzed within 3-5 minutes, as longer incubation will increase blue colored cells. Trypan blue staining may result in an overestimate of viable cells especially for low viability samples.

  6. Explant culture

  7. Chicken embryo culture for virus cultivation

  8. Cytotoxicity assays 1-Neutral red uptake 2- MTT 3- Crystal violet Neutral red uptake

  9. Principle: Formazan crystals are formed by mitochondrial succinate dehydrogenase in living and early apoptotic cells, but not in dead cells. MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide) MTT cytotoxicity test in cell culture http://www.ib.amwaw.edu.pl/home/dslado/video/mtt.html

  10. Umbilical cord cell culture

  11. Cell Identification Flowcytometry Immunohistochemistry

  12. Improvement cell culture research Establish suitable laboratories for cell culture. Supply continuous electricity.

  13. Importing cells, reagents and growth factors in suitable cooled containers. Establish cell bank for continuous cell supply.

  14. Future work Production of monoclonal antibodies Tissue Engineering

  15. Growing cells in 3D Forming Tissue in Bioreactor stem cell-mediated therapy

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