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