Download - Tissue Culture Technology,242013
-
7/28/2019 Tissue Culture Technology,242013
1/63
ANIMALTISSUE CULTURE
-
7/28/2019 Tissue Culture Technology,242013
2/63
Topics
Introduction1
Tissue Culture2
Hybridoma Tech3
Future Medicine4
-
7/28/2019 Tissue Culture Technology,242013
3/63
-
7/28/2019 Tissue Culture Technology,242013
4/63
Biotechnology: Present and Future
-
7/28/2019 Tissue Culture Technology,242013
5/63
-
7/28/2019 Tissue Culture Technology,242013
6/63
IMMUNITY
-
7/28/2019 Tissue Culture Technology,242013
7/63
Figure 1-7Anatomi
sistemimun
-
7/28/2019 Tissue Culture Technology,242013
8/63
CONTROL OF VIRUS INFECTION
-
7/28/2019 Tissue Culture Technology,242013
9/63
T Helper (Th) Cells in Immune Responses
Th cell
Cellular
Immunity
Humoral
Immunity
Th cells-Play a central role in immune protection.
-Orchestrate immune responses through
their production of cytokines and
chemokines.
-
7/28/2019 Tissue Culture Technology,242013
10/63
Th cell subsets
Th1Th17
TregTh2 Th cell
-
7/28/2019 Tissue Culture Technology,242013
11/63
Diseases with Imbalance of
Th1/Th2 Response
Th1 > > Th2 Th1 < < Th2
VS
Autoimmune diseases
Multiple sclerosis
DiabetesRheumatis arthritis
Viruses
HIV, MAIDS
HSV-1
BacteriaLister ia
monocytogenesOrgan transplantation
Allergic diseases
Allergy
AsthmaAllergic rhinitis
ProtozoansLeishmania major
Tryanosoma cruzi
BacteriaMycobacter ium
tuberculosisFungiCandida
-
7/28/2019 Tissue Culture Technology,242013
12/63
Figure A-35Cel l St imu lat ing Agen ts
-
7/28/2019 Tissue Culture Technology,242013
13/63
-
7/28/2019 Tissue Culture Technology,242013
14/63
Future Medicine
-
7/28/2019 Tissue Culture Technology,242013
15/63
Epidemiology
Surveillance &
Disease Investigation
Biostatistics
Environmental
Science
Community Health
Promotion
Infectious Disease
Competencies
-
7/28/2019 Tissue Culture Technology,242013
16/63
Morens, D., Folkers, G., & Fauci, A.(2004) The challenge of emerging and re-emerging infectiousdiseases. Nature, 430: 242-249.
-
7/28/2019 Tissue Culture Technology,242013
17/63
Global distribution of Relative Risk for an EID Event
caused by Zoonotic Pathogens from Wildlife*
Jones KE, Patel NG, Levy MA et al. Nature 2008; 451:990-993.
-
7/28/2019 Tissue Culture Technology,242013
18/63
-
7/28/2019 Tissue Culture Technology,242013
19/63
-
7/28/2019 Tissue Culture Technology,242013
20/63
Inhibition of IL-12 Production as a
Therapeutic Strategy of Autoimmune diseases
To screen active compounds with inhibitory
activity on IL-12 production1. Retinoids, oxLDL
2. Chemicals from some medicinal plants
3. Combinatorially synthesized chemicals
Major IL-12 producers
Macrophage Dendritic cell
-
7/28/2019 Tissue Culture Technology,242013
21/63
Tissue Culture: Teory, Techniques,and Application
D i d
-
7/28/2019 Tissue Culture Technology,242013
22/63
Design dan Layout
Laboratorium
Service Bench
Preparation room
Wash up (Tempat pencucian)
Storage
Construction and layoutEssential Equipment
-
7/28/2019 Tissue Culture Technology,242013
23/63
-
7/28/2019 Tissue Culture Technology,242013
24/63
Sterile Handling Area
Laminar flow
Incubation
Heaters
Air circulation
Thermostats
-
7/28/2019 Tissue Culture Technology,242013
25/63
-
7/28/2019 Tissue Culture Technology,242013
26/63
-
7/28/2019 Tissue Culture Technology,242013
27/63
-
7/28/2019 Tissue Culture Technology,242013
28/63
-
7/28/2019 Tissue Culture Technology,242013
29/63
-
7/28/2019 Tissue Culture Technology,242013
30/63
PENANGANAN KONTAMINASI
Sumber infeksi
1. Sel
2. Media kultur
3. Glassware dan Plastic ware
4. Airborne modes
Laminar flow TransferRoom
-
7/28/2019 Tissue Culture Technology,242013
31/63
-
7/28/2019 Tissue Culture Technology,242013
32/63
-
7/28/2019 Tissue Culture Technology,242013
33/63
Biologi cell culture
Kuliah ke 3
-
7/28/2019 Tissue Culture Technology,242013
34/63
Petunjuk praktis kultur sel
Ruang kultur dan lingkungannya
Sterilitas dan peralatan
Kualitas bahan untuk kulturPemilihan Media
Pemilihan Serum
Material untuk kultur sel
-
7/28/2019 Tissue Culture Technology,242013
35/63
Biologi Sel Kultur
Lingkungan kultur: In vivo x in vitro
2 16
Transformation
Cell death
Log cell
-
7/28/2019 Tissue Culture Technology,242013
36/63
Istilah
Dedifferentiation:
Undefferentiated cell
Absence of appropriate inducer
Penting untuk di pahami: dedifferentiation,deadaptation, differentiation and selection
CLONE SELECTION THEORY
-
7/28/2019 Tissue Culture Technology,242013
37/63
CLONE SELECTION THEORY
-
7/28/2019 Tissue Culture Technology,242013
38/63
-
7/28/2019 Tissue Culture Technology,242013
39/63
-
7/28/2019 Tissue Culture Technology,242013
40/63
PENANGANAN KONTAMINASI
Sumber infeksi
1. Sel
2. Media kultur
3. Glassware dan Plastic ware
4. Airborne modes
Laminar flow TransferRoom
-
7/28/2019 Tissue Culture Technology,242013
41/63
PENANGANAN KONTAMINASI
Teknik Aseptik
Antibiotika
Prosedur di Ruang Transfer
P d di R T f
-
7/28/2019 Tissue Culture Technology,242013
42/63
Prosedur di Ruang Transfer
Personel Ruang kultur
Bahan yang dipakai harus di tes
sebelumnya (media, serum, trypsin,
HBSS dan komponen lainnya dari
media)
KulturSkills
-
7/28/2019 Tissue Culture Technology,242013
43/63
TC APPLICATION
T Helper (Th) Cells in Immune Responses
-
7/28/2019 Tissue Culture Technology,242013
44/63
T Helper (Th) Cells in Immune Responses
Th cell
Cellular
Immunity
Humoral
Immunity
Th cells-Play a central role in immune protection.
-Orchestrate immune responses through
their production of cytokines and
chemokines.
-
7/28/2019 Tissue Culture Technology,242013
45/63
-
7/28/2019 Tissue Culture Technology,242013
46/63
Teknik Hibridoma
o Isolasi Limfosit
o Isolasi Makrofag
o Kultivasi Myeloma
o Fusi Sel
o Seleksi Hibrid
-
7/28/2019 Tissue Culture Technology,242013
47/63
HAT-SELECTION
B-cells Non fused B-cells
Non fused myeloma
Fusion
False hybrid
Myeloma cells True Hybrid
-
7/28/2019 Tissue Culture Technology,242013
48/63
Hybridoma clones and normal feeder cells
THE STEP THAT YOU SHOULD FREEZE
-
7/28/2019 Tissue Culture Technology,242013
49/63
THE STEP THAT YOU SHOULD FREEZE
STOCK IN HYBRIDOMA PRODUCTION
Growing myeloma cells from freeze stock
fusionScreen
Split pos clones into 24 well plates
Screen to weed out false pos
split pos clones into T-25 flasks
Freeze at least Single cell clone2 vials/ clone
Screen
Split pos clones into plates
split in to culture flasks
Freeze stock
Expanded for Ab production
Ad t f Ab d Ab
-
7/28/2019 Tissue Culture Technology,242013
50/63
Advantage of mAb and pAb
mAb pAb
Homogeneous of product Heterogeneous of productonly one specificity Multiple specificity
constant Reactivity Variable reactivity
Unlimited Limited
No diff. batch-to batch Batch-to batch variationImmunization with crude antigen Immunization with purified antigen
Small doses for therapy High doses for therapy
Di d t f Ab d Ab
-
7/28/2019 Tissue Culture Technology,242013
51/63
Disadvantage of mAb and pAb
mAb pAb
High production cost Low production cost
Long of production: 4-6 months 3-6 monthsDefinite affinity & Ig class Variable
Definite biol. Character & funct. Variability
No agglutination reaction Agglutination
reaction
-
7/28/2019 Tissue Culture Technology,242013
52/63
Figure A-15Antibody engineering
-
7/28/2019 Tissue Culture Technology,242013
53/63
Figure A-15 part 2 of 2
-
7/28/2019 Tissue Culture Technology,242013
54/63
Diferensiasi sel
Dedifferentiation:
Undefferentiated cell
Absence of appropriate inducer
Penting untuk di pahami: dedifferentiation,
deadaptation, differentiation and selection
To oplasma gond i i
-
7/28/2019 Tissue Culture Technology,242013
55/63
Toxoplasma gond i i
-
7/28/2019 Tissue Culture Technology,242013
56/63
-
7/28/2019 Tissue Culture Technology,242013
57/63
Aplikasi TC
-
7/28/2019 Tissue Culture Technology,242013
58/63
Virus propagation
Trypanosoma
-
7/28/2019 Tissue Culture Technology,242013
59/63
Trypanosoma
INVASI T gond ii
-
7/28/2019 Tissue Culture Technology,242013
60/63
INVASI T. gond ii
Apoptosis
-
7/28/2019 Tissue Culture Technology,242013
61/63
Apoptosis
-
7/28/2019 Tissue Culture Technology,242013
62/63
Genetic disorders:Kl inefel ter sy ndrom e
-
7/28/2019 Tissue Culture Technology,242013
63/63
Life long learner & building net work,
TERIMAKASIH