Transcript
Advanced Aminal Cell Culture
2013 2nd Semester
Department of Animal Science
Chungbuk National University
5rd Lecture
Syllabus
Date TopicsSeptember 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 INovember 14, 2013 Stem Cell IINovember 28, 2013 TG/KO Animals December 5, 2013 Genome Engineering/NGS
December 12, 2013 Final Exam
DateSeptember 26, 2013 Cell Culture For
Antibody / Protein Production
조유미 , Madhusumida
October 17, 2013 Protein Production/Purification 이미진 , 정용호
October 31, 2013 이영 , 윤준호November 14, 2013 Stem Cell I Jia Jia Lin, 염동현
November 28, 2013 Stem Cell II Zhao MingHui,권정우December 5, 2013 Transgenic Animals Lin Zili, 이상배December 12, 2013 Genome
Engineering/NGS 조유진 , 김상욱
What is Stem Cell?
A primitive cell which can either self renew (reproduce itself) or give rise to more specialised cell types
Stem cell is the ancestor at the top of the family tree of related cell types,
e.g.) one blood stem cell gives rise to red cells, white cells and platelets
Characteristics of Stem Cell
Totipotent : can give rise to all types of adult tissue cells
plus extraembryonic tissue. can give rise to a new individual
given appropriate maternal support
Pluripotent : descendants of totipotent cells and can
differentiate into nearly all cells. cells which support
embryonic development
Multipotent : can give rise to several types of
mature cell, but only those of closely related family.
Self Renewal
- Stem cell can self-renewal by itself indefinetly
- Balance between differentiation and Self-Renewal should be maintained.
Symmetric cell division
Asymmetric cell division
1. Self-renews 2. Differentiates
Progenitor cell
Stem cell Stem cell
SELF – RENEWAL
DIFFERENTIATION
DIFFERENTIATION
Embryonic stem cells
Derived from embryos before specialised tissues of the body begin to form (epiblast)
- grown indefinitely in culture in the primitive embryonic state
- Retain the property of pluripotency during extended growth in vitro
Cell Lineage
Maintenance of ESC
- Special medium are required to maintain pluripotency- Leukemia inhibitory factor (LIF) : Removal of LIF pushes stem cells toward diffrentation- LIF is essential for mouse embryonic stem cell culture
Human ES cell maintenance
Culture condition: LIF-independent
Serum: FGF, nodal/activin and Wnt and KSR
mESC hESC
Difficulties in culture Relatively easy Relatively difficult
Colony shape Dome-like structure Flatten structure
Growth rate Relatively fast slow
Coating plate Not required Matrigel
Passage Trypsinization Mechanical passage
Media requirement LIF, mouse embryonic fibroblast
No LIF required, bFGF, Growth with/without feeder
Seperation Easy, No regulation Difficult, Strict Regulation, Ethical Problem
Moleclar Basis of Pluripotency and Cell Differentiation
Histone Modification
Molecular Basis of Cell Differentation
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