ANIMAL CELL TECHNOLOGY

Lecturer: Nguyen Thi Hong Minh

Email: nguyen-thi-hong.minh@usth.edu.vn
Tel. 094 393 6511
 Overview

 L1: Introduction to animal cell technology

 L2: Basic method in animal cell culture

 L3: Animal cell as tool for drug discovery

 L4: Animal cell as tool for biologist

 L5: 2D/3D culture, antibody production

 L6: Good laboratory practice (GLP) and good cell culture practice

 L7: Student presentation

Cell – cell interaction and
communication
Organization of Life
 Cell- cell Interaction

 There are 2 types: homotypic and

heterotypic.
Cell adhesion can occur homotypically (i.e.
between the same cell types) or
heterotypically (i.e. between two different
cell types)

 Cells can identify each other by cell surface markers.

 glycolipids are commonly used as tissue- specific markers
 major histocompatibility complex (MHC) proteins are used by
cells to distinguish “self” from “non-self”
 Extracellular matrix

- Most animal cells release

materials into the extracellular
space
these materials are called
the extracellular matrix
- Extracellular matrix fills the
spaces between cells and binds
cells and tissues together

 the extracellular matrix not only holds the cells together to form a tissue, but
also allows the cells within the tissue to communicate with each other
 Animal cell contacts include tight junctions, gap junctions,
and desmosomes

 Plasmodesmata are junctions between plant cells

 Tight junctions

 A tight junction is a
watertight seal between two
adjacent animal cells
 The cells are held tightly
against each other by proteins
 This tight adherence prevents
materials from leaking
between the cells
 Ex: the tight junctions of the
epithelial cells lining your
urinary bladder prevent urine
from leaking out into the
extracellular space
Tight junctions
 Desmosomes

 Act like spot welds

between adjacent epithelial
cells
 Short proteins called
cadherins in the plasma
membrane connect to
intermediate filaments to
create desmosomes
 The cadherins join two
adjacent cells together and
maintain the cells in a
sheet-like formation in
organs and tissues that
stretch, like the skin, heart,
and muscles
 Gap Junctions

 they are channels

between adjacent
cells that allow for
the transport of ions,
nutrients, and other
substances that
enable cells to
communicate
 formed by two
unrelated protein
families, the
pannexins
and connexins
 Gap Junctions

 Gap junctions are

particularly important in
cardiac muscle: The
electrical signal for the
muscle to contract is
passed efficiently through
gap junctions, allowing the
heart muscle cells to
contract in tandem

 When the pores (“doughnut

holes”) of connexons in
adjacent animal cells align, a
channel between the two cells
forms
 Cell communication

 Chemical signals (hormones, etc) travel

throughout animals (plants) to target cells and
convey information from one tissue or organ
to another. This intercellular signaling involves
four steps: signal reception, signal processing,
signal response, and signal deactivation

 The function and chemical structure of animal

(plant) hormones vary widely. However, all
hormones play a role in coordinating cell
activity in response to information from
outside or inside the body
 Reception: a signal molecule
binds to a reception or protein,
causing it to change shape

 The binding between a signal

molecule (ligand) and a
receptor is highly specific
 Intracellular receptors: inside
plasma membrane in the
cytoplasm or nucleus.
 Plasma membrane receptors:
bind to water-soluble ligands
Signal response and deactivation

 Signal response include changes in gene expression

and changes in the activity of specific proteins

 Turning off cell signals is just as important as turning

them on. Cells have automatic and rapid mechanisms
for signal deactivation. These mechanisms allow the
cell to remain sensitive to future hormone signals
cell communication
 Cell communication

 Cell Communication: Direct contact

– molecules on the surface of one
cell are recognized by receptors on
the adjacent cell
 Cell communication

 Cell Communication: Paracrine

signaling – signal released from a
cell has an effect on neighboring
cells
 Cell communication

 Cell Communication: Endocrine

signaling – hormones released from
a cell affect other cells throughout
the body
 Cell communication

 Cell Communication: Synaptic signaling

– nerve cells release the signal
(neurotransmitter) which binds to
receptors on nearby cells
Animal cell as tool for drug discovery

o Non-targeting drug discovery

Cytotoxicity assay
Apoptosis assay

o Targeting drug discovery

Reporter assay
 What is cancer?
 Caner is defined as the continuous uncontrolled growth of cells.
Loss of normal growth control

Normal cell
division

Cell suicide or apoptosis

Cell damage with no repair
MTT assay
Necrosis & Apoptosis
Annexin V staining
Flow cytometry
Analyse cell apoptosis by flow
cytometry
Animal cell as tool for drug discovery

o Non-targeting drug discovery

Cytotoxicity assay
Apoptosis assay

o Targeting drug discovery

Reporter assay