Swan(sea) Song – personal research during my six years at Swansea ... and bey...
Cell culture 05
1. 5rd 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
4. 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
5.
6. 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.
7.
8.
9.
10.
11. Self Renewal
- Stem cell can self-renewal by itself indefinetly
- Balance between differentiation and Self-Renewal
should be maintained.
16. 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
19. 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
20.
21. Human ES cell maintenance
Culture condition: LIF-independent
Serum: FGF, nodal/activin and Wnt and KSR
22. 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
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!
Then at blastocyst formation and gastrulation the stem cells start to divide asymmetrically. When these stem cells divide, they give rise to two cells that are different from each other. One of the cells remains a stem cell—the yellow one—and the other changes into a progenitor cell—the green square. That progenitor cell is a young cell that will change into a mature cell type, like the epithelial cell in green.Asymmetric cell division is when a stem cell divides to produce two cells that are different from each other. CLICK! One of the cells remains a stem cell, demonstrating self-renewal, and the other CLICK! differentiates into a progenitor cell.
Ok so that stem cell divided to produce a green progenitor cell which differentiates into a green skin cell (CLICK!). That stem cell can divide again,this time producing a different type of progenitor cell CLICK! which matures into yet another cell type, CLICK! like this brain cell.OK so what makes a stem cell unique? (wait for answers) FIRST: it maintains its own population—self-renewal—and SECOND: its capacity to give rise to different progenitors that change into a mature cell—differentiation.
