Nhap mon cong nghe sinh hoc

Animal culture
Kỹ thuật nuôi cấy tế bào động vật
Cell culture


Tissue
Tripsin
Suspended cell/medium
Adhere to bottom of a flask and form a monolayer
Proliferation then stop
subculture
Treat with EDTA/tripsin
(take out from flask)
Secondary culture
Group of cell strain
t0
proliferation
2 main categories of animal cell cultures….
Primary culture

Continuous cell line
Primary Cultures
Taken from fresh tissue

Limited life span in culture

Treated by proteolytic enzyme (Trypsin)

Separate into single cells
-epithelial cells
-fibroblasts
Continous cell lines
Not any primary cell and its generation are die after proliferating.However, some cells continue growing. They have different morphology, grow faster. Their generation exist unlimited –called cell line
Through transfer subculture they may be changed their property. EX: increasing group of cell irregularity (neoplastic---tế bào ung thư)
Cell + glycerin, dimethylsulphosid kept Cool preservation to prevent cold crystal then placed into LN
Continuous Cell Line
Derived from humans

Been transformed
-lose sensitivity to factors associated with growth control

Produce immortalised cell lines

Cell lines are neoplastic

Often lose their anchorage-dependence
-associated with an altered xsome pattern

More easily cultured
Medium
Enough nutrition for cell growing
Ph = 7.0-7.3 (need buffer)
Same isotonic with cytoplasm
Sterility
It should have :
Inorganic ion
Precise osmotic
Ph
Energy (glucose)
pH Indicator (phenol)
AA (Arg, Cys, Tryp, tyr,Meth, Glu, Isoleu, Leu, Lys, phe,…)
Vit (acid Floric, Inositol, Nicotinic, B6, B2, B3…)
Buffer (Bicarbonatnatri)
Antibiotic
Serum (5-10%)
Water ion- free

Standard for cultural medium
Application of animal cell culture
In vitro toxicity testing
Produce virus vaccines
Chicken embryo/ cell culture
Animal cell culture
(kidney of monkey or checken embryo)
Infection with virus
Collect virus after replication
Purification virus by filter/condense
Suspended virus +stably material to prevent decease activity – keep under cold condition
Produce Hormones
What Are Stem Cells?
Stem cells are the raw material from which all of the body’s mature, differentiated cells are made. Stem cells give rise to brain cells, nerve cells, heart cells, pancreatic cells, etc.

How do cells remain
Self renewing and
unspecialized?
What are the signal????
Hormones
Growth factors
Small proteins

Definition: is able to proliferation continuously and develop into differentiation cell
Stem cell
Totipotency –The capacity of a cell (or a group of cells) to give rise to an entire organism.
Germ cell: capacity to divide into one or definite special cell.
EX: blood germ cell develop into different blood cells (red blood, leukocyte, lymphocyte)


STEM CELL BASICS
Where do stem cells come from?
Sources of stem cells during development
Teratomas” (benign);
“Teratocarcinomas”
(malignant
[P] = pluripotent
[M] = multipotent
[T] = totipotent
Origin of Human Embryonic Stem Cells

Isolation of Human Embryonic Stem Cells
First done by Dr.
James Thompson
and colleagues at
the University of
Wisconsin (1998
Properties of Human Embryonic Stem Cells in culture

• Pluripotent – able to form any of ~200 different types of cells of the body
• Self renewing in vitro – can propagate or proliferate indefinitely in the undifferentiated state
• Express the enzyme telomerase (required to maintain the ends of chromosomes) and Oct4 (a master regulator of ESC pluripotency)
• Maintain normal chromosome structure and complement even after long periods in culture (unlike many other tissue culture cell lines)
What’s So Special About Stem Cells?
They have the potential to replace cell tissue that has been damaged or destroyed by severe illnesses.

They can replicate themselves over and over for a very long time.

Understanding how stem cells develop into healthy and diseased cells will assist the search for cures.
Two Kinds of Stem Cells
Embryonic (also called “pluripotent”) stem cells are capable of developing into all the cell types of the body.

Adult stem cells aer less versatile and more difficult to identify, isolate, and purify.
Embryonic Stem Cells
Researchers extract stem cells from a 5-7 days old blastocyst.

Stem cells can divide in culture to form more of their own kind,
thereby creating a stem cell line.
The research aims to induce these cells to generate healthy tissue needed by patients.
Two Kinds of Stem Cells
Embryonic (also called “pluripotent”) stem cells are capable of developing into all the cell types of the body.

Adult stem cells are less versatile and more difficult to identify, isolate, and purify.
Two Sources of
Embryonic Stem Cells
1 Excess fertilized eggs from IVF (in-vitro fertilization) clinics

2. Therapeutic cloning (somatic cell nuclear transfer)
Tens of thousands of frozen embryos are routinely destroyed when couples finish their treatment.
These surplus embryos can be used to produce stem cells.
Regenerative medical research aims to develop these cells into new, healthy tissue to heal severe illnesses.
Somatic Cell Nuclear Transfer
The nucleus of a donated egg is removed and replaced with the nucleus of a mature, "somatic cell" (a skin cell, for example).
No sperm is involved in this process, and no embryo is created to be implanted in a woman’s womb.
The resulting stem cells can potentially develop into specialized cells that are useful for treating severe illnesses.
Why do it?
Particular cells can be isolated and cloned
-isolation of mutant cells
-investigate cell growth
-produce hybrid cells that have applications in biotechnology

Produce important pharmaceuticals
-vaccines
-hormones
Stem cell cultivation
Fertilized egg
Blastocyst
Cultured undifferentiated
stem cells
Differentiation
Blood cells
Muscle
Neural cells
Embryo
Isolated
pluripotent
stem cell
Origin of Human Embryonic Stem Cells
Isolation of Human Embryonic Stem Cells
First done by Dr.
James Thompson
and colleagues at
the University of
Wisconsin (1998).
Application of stem cells


1. Stem cells used for pharmaceutical safety
2. Cell therapy
Embryo transfer