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Cell culture 06
1. 6th Lecture
Advanced Aminal Cell 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
Cell Culture 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
5. Differentiated Cell vs Stem 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 Cell Differentitation
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?
11. How to return back 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)
12.
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 John Gurdon (1962)
Conclusion : Oocyte have ability to reprogramming terminally differentiated cell
20. Drawbacks of Stem Cell Generation using SCNT
- Ethical Problems
• ES cell should be generated from Cloned Blastocyst
• Human Oocyte is needed
• Human Cloning?
- Technical Problems : Low efficiency
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
- Retroviral vector : 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?
30.
31. ‘OSKM’ is not enough? Presence of ‘Break’
Rais et al., Nature 2013
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!
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!
