Kingsborough Community College of The City University of New YorkDepartment of Biological Sciences
Biology 57 – Biotechnology: Cell Culture and Cloning
BIO 05700 – BIOTECHNOLOGY: CELL CULTURE AND CLONING (4 crs. 6 hrs.(2hrs lecture, 1hr recitation, 3 hrs laboratory))Biological principles underlying animal and plant tissue culture and cloning techniques are taught using current cell culture research publications. The formation and maintenance of primary and continuous culture, monolayer and suspension cultures, cell separation techniques, and cell cloning studies are conducted. Cell morphology in vitro, callus formation, cytochemistry, immunocytochemistry, autoradiography, chromosome spread preparation and karyotyping western blotting and enzyme-linked immunoabsorbance assays are performed and analyzed. Photomicroscopy and independent research projects on the initiation and maintenance of primary cell cultures from such sources as chick embryos are also performed.Prerequisites: BIO 01400, CHM 01100 or Department Permission
Topical Course Outline
Text Book: Freshney, R.I. 2005. Culture of Animal Cells 5th ed. Wiley-Liss. Pp 642. Lab Manuals: 1. Freshney, R.I. 2005. Culture of Animal Cells 5th ed. 2005. Wiley-Liss. Pp 642. 2. Laimer, M., and W. Rücker. eds. 2003. Plant Tissue Culture. Springer. Pp 260.
Introduction to biology of cells in culture. Design layouts for a cell culture laboratory: similarities and differences for plant and animal cell culture.
Laboratory Safety and Biohazards. Practice of aseptic techniques for sterilization of apparatus, reagents and media.
Special environment for cell culture - substrate, gas, medium and temperature for animal and plant cell culture.
Disaggregation of animal tissue and primary culture. Begin plant cell callus and suspension cultures.
Methods of cell separation: by cell size, sedimentation velocity, isopychnic sedimentation, flourescence-activated cell sorting and others.
Routine maintenance of animal cell line, and subculture. Continue plant cell callus and suspension culture.
1st Lecture Exam. Cell cloning and cell selection: dilution cloning, semisolid media cloning, cloning in methocel over agar and others. Induction of cell differentiation in culture.
1st quiz. Animal cell cloning and cell selection. Continue plant cell callus culture until organogenesis.
The Transformed Phenotypes of transformed animal cells – transformation, anchorage, genetic abnormalities and invasiveness.
Induction differentiation of animal cells. Isolation and culture of plant cell protoplasts.
Characterization of animal cells using morphology, chromosome, DNA, RNA and protein content.
Study differentiated animal cell phenotypes. Examine and continue plant protoplast culture.
Animal and plant cell culture contamination, instability, variation, and preservation and cell quantitation.
Quantitation of animal cell growth.Examine and continue plant protoplast culture.
2nd Lecture Exam.Plant cell transformation methods. Gene gun particle bombardment.
2nd quiz.Experiments on animal cell viability and cytotoxicity.Cross species plant cell fusion culture.
Specialized animal cell culture: Tumor tissue, epithelial, mesenchymal, neuroectodermal and hemopoietic cells.
Determine animal cell viability from the previous culture.Begin animal special cell culture.Cross species plant cell fusion culture.
Three-dimensional animal cell culture systems for organ culture, histotypic culture, and filter wells
Continue special animal cell culture. Continue growing fused plant cells.
Specialized animal cell culture techniques – mass culture, lymphocyte preparation, isect and cell culture, poikilotherm cell culture and others.
Set up mass culture of HeLa cells.Document growth, morphology and stability characteristics of fused plant cells.
Virus preparation and assay.Pigment production from cell culture.
Record and analyze results of mass animal cell culture.Analyze the results from fused plant cell culture. 3rd quiz.
Final Exam Cumulative.