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Cell culture 06
This document contains the syllabus for an advanced animal cell culture course from the Department of Animal Science at Chungbuk National University. The syllabus lists 8 topics that will be covered between September 2013 and December 2013, including introductions to cell culture and stem cells, using cell culture for antibody and protein production, transgenic and knockout animals, and genome engineering. Student groups are assigned to present on specific dates, including topics like stem cell I and II and protein production/purification. The document also includes an excerpt from the November 14th lecture on stem cell II, discussing the mechanisms of cellular reprogramming and how induced pluripotent stem cells were discovered.
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Cell culture 06
- 1. 6th Lecture Advanced AminalCell Culture 2013 2nd Semester Department of Animal Science Chungbuk National University
- 2. Syllabus Date Topics September 5, 2013 Introduction: What is Cell Culture? September 12, 2013 Cell Culture As Model System For Research September 26, 2013 Cell Culture For Antibody / Protein Production October 17, 2013 Protein Production/Purification October 31, 2013 Stem Cell I November 14, 2013 Stem Cell II November 28, 2013 TG/KO Animals December 5, 2013 Genome Engineering/NGS December 12, 2013 Final Exam
- 3. Date September 26, 2013 CellCulture For Antibody / Protein Production 조유미, Madhusumida October 17, 2013 Protein Production/Purification 이미진, 정용호 이영, 윤준호 October 31, 2013 November 14, 2013 Stem Cell I Jia Jia Lin, 염동현 November 28, 2013 Stem Cell II Zhao MingHui,권정우 December 5, 2013 Transgenic Animals Lin Zili, 이상배 December 12, 2013 Genome Engineering/NGS 조유진, 김상욱
- 4. Stem Cell II Today’stopic : Reprogramming
- 5. Differentiated Cell vsStem Cell Cell Differentation The process by which cells or parts of an organism become different from one and other and also from their previous state. The process by which cells or tissues of an organism acquire the ability to perform their special functions.
- 6. Mechnism of CellDifferentitation How is this accomplished? Selective gene transcription In any given cell, only the genes necessary for basic metabolism and that cell’s special functions are active. * muscle cells - actin, myosin * pancreatic acinar cells - digestive enzymes * neurons – neurotransmitters How selective gene transcription could be achieved?
- 7. Changes in epigeneticmarks during Development Messiner, Nature Biotechnol. 2010
- 9. Bock et al.,Mol.Cell 2012
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- 11. How to returnback to pluripotent state? Things to do : - Reset all of epigenetic marks which repress pluripotent factors maintenance genes (Activation) - Reset all of epigenetic marks which activate lineage specific gene expression (Repression)
- 13. Natural Reprogramming :Sperm & Oocyte Sperm is highly differentated Cell. After fertillization, DNA derived from sperm is demethylated rapidly Smith et al., Nature 2012
- 14. Experiments by JohnGurdon (1962) Conclusion : Oocyte have ability to reprogramming terminally differentiated cell
- 15. Somatic Cell NuclearTransfer (SCNT)
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- 19. Stem Cell fromSomatic Cell Nuclear Transfer (SCNT)
- 20. Drawbacks of StemCell Generation using SCNT - Ethical Problems • ES cell should be generated from Cloned Blastocyst • Human Oocyte is needed • Human Cloning? - Technical Problems : Low efficiency
- 21. iPSC (induced PluripotentStem Cell)
- 22. ‘Yamanaka Factor’ - Oct4,Sox2, Klf-4, c-Myc (OSKM) - Transcription Factors which express abundantly in Embryonic Stem Cell - Screened from 24 transcription factors expressed in ESC - Retroviral expression of these 4 genes in embryonic/Adult fibroblast transform Cells into ‘Stem Cell Like’ cells
- 23. Molecular event ofinduced pluripotency
- 27. Cell Lineage SpecificGene Pluripotency Maintenance genes
- 28. Limitation of iPSC -Although iPSC is much similar with ESC, its properties is not exactly same with those of ESC • Source of Original Cell • Condition used for the generation of iPSC Different epigenetic states Different tumorgenicity … - Potential Tumorgencity of iPSC
- 29. Methodologic Limitation - Retroviralvector : Danger in the tumor inducing - Known oncogene (c-Myc) was one of component - Low efficiency for reprogramming Efforts to overcome these Limitation - Less factors (Sometime Only Oct4 can induce pluripotency) -Protein-mediated generation of iPSC - Chemical mediated?
- 31. ‘OSKM’ is notenough? Presence of ‘Break’ Rais et al., Nature 2013
- 32. Current Application ofiPSC - Research Tools : Patient specific cell derived from iPSC
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- 38. High-throughput screening ofdrug candidate using iPSC derived cells
Editor's Notes
- #16 At the beginning of embryonic development, stem cells undergo symmetric cell division. They divide symmetrically, where one cell splits and gives rise to two identical cells that have the same potential. This is why we said the stem cells in the early embryo remain the same and are identical. Interestingly, this is also how fully mature cells in your body divide. What’s the technical term for this that you guys probably learned in biology? (wait) MITOSIS!
- #22 At the beginning of embryonic development, stem cells undergo symmetric cell division. They divide symmetrically, where one cell splits and gives rise to two identical cells that have the same potential. This is why we said the stem cells in the early embryo remain the same and are identical. Interestingly, this is also how fully mature cells in your body divide. What’s the technical term for this that you guys probably learned in biology? (wait) MITOSIS!