7/28/2019 CTS StemCell Lecture
1/8
1
Week 8
STEM CELLS
Carolin e Tan Sardjono , dr., PhD
Fundamental Medical Science I
2009
Stem Cell Lecture (6 x 50 minutes)
To understand:1. Stem cell characteristics
Lecture 1: Stem Cell in clinical setting
Duration 3 x 50 minutesWeekly objectives:
. em ce ypes3. Stem cell sources4. Stem cell mechanisms of action5. Stem cell for clinical application6. Immunological basic knowledge for stem cell
transplantation7. Basic knowledge in stem cell banking for clinical
use
Stem Cell Lecture (6 x 50 minutes)
Lecture 2: Advances in Stem Cell ResearchDuration 3 x 50 minutes
Weekly objectives:
1. Alternative sources for stem cell isolation
2. Embryonic stem cell (In vitro Fertilization,Somatic Cell Nuclear Transfer, Parthenogenesis)3. Induced pluripotentstem cell (iPS)4. Stem cell application in clinical trials
Stem Cell Overview
Living Organisms need to be able to survive manychallenges over their life
Survival should be able to regenerate injured &damages cells
Skin, blood in circulation, intestinal mucosa,epithelial cells in respiratory tract, all kind of cellsin our body need to regenerate
The regeneration process is roled by thestem cells
Stem Cell Overview
Recently, stem cells have been considered as apotential therapy to treat many diseases includingmalignancy, hematological abnormalities, anddegenerativediseases
Moreover, stem cells can be- stored in the longer term for future usage,- ex-vivo expanded,- differentiated, and- generated into specific organs.
7/28/2019 CTS StemCell Lecture
2/8
2
Video stemcell1.mp4
A rather nice illustration about stem cells
Covers briefly about embryonic stem cells (tommorows
lecture) but good intro about adult stem cell
Stem Cell Characteristics
1. Self renewal (divide indefinitely)
2. Undifferentiated (unspecialized)3. Ability rise other cell types
STEM CELL
Source: NIH booklet
Undifferentiated
Cell
Different kinds of Stem Cell
I. Embryonic Stem Cell (lecture 2)
II. Adult Stem Cell
Cultured in vitro
lipoaspirate Apheres isBone marrow
Umbilical cord
blood
Adult Stem Cell
1. Hemato oeticStem Cell CD34+
2. Mesenchymal Stem Cell
1st type of stem cell
Hematopoetic
Stem Cell (CD34+)
GenerateBlood Cells
MononucleatedCells with
1. Hematopoetic Stem Cell
In our Body:Reside in Bone Marrow
0.01-0.1% present in blood circulation
CD34
CD34
7/28/2019 CTS StemCell Lecture
3/8
3
A population known as Hemangioblast+ +
1. Hematopoetic Stem Cell
A proportion of HematopoeticStem Cell
can also raise endothelial cells forblood vessels regeneration
Able to differentiateinto EndothelialProgenitor CellsMature asEndothelial cells
2nd type of stem cell
Mesenchymal
Stem Cell
Also known as CD34- Stem CellIn our Body:1. Present as solid structure in Bone Marrow2. Within sub cutaneous fat, embedded among our fat
cells
2. Mesenchymal Stem Cell
fibroblast-like
structure
100m100m
Criteria to define MSC
2. Mesenchymal Stem Cell
1. Bone marrowAdult Stem Cell Sources Adult Stem Cell Sources
2. Peripheral Blood
Blood collection through arm
vein directly to blood bag
Blood collection through arm vein
post-concentrated by
apheresis machine
For high cell number use Granulocyte Colony Stimulating
Factor tomobilizethe stem cells from Bone Marrow to blood
circulation
7/28/2019 CTS StemCell Lecture
4/8
4
Adult Stem Cell Sources
Collected fromumbilical vein
3. Cord Blood Whycollectedfrom
Adult Stem Cell Sources
Cord Blood collection
After the baby wasborn
Collection fromumbilical vein atthe placentas part
Lipoaspirate
Adult Stem Cell Sources
Following liposucction procedure
Good source for mesenchymal stem cellUsed in reconstructive surgery
Adult Stem Cell SourcesOther Sources (low cell yield, for research) :1. Liver2. Hair follicle3. Endometrium.
5. Etc...
1 2 4
How Stem Cell works
1.To Restore hematopoiesis following highdose chemotherapy or radiation and toreplace defective cells (e.g. leukemic cells,autoreactive cells in autoimmune diseases)
Before the stem cells repopulate the immune cells,patient is required to stay in the isolation room
Have been done since 1970s
7/28/2019 CTS StemCell Lecture
5/8
5
How Stem Cell works2. In reconstructive surgery:
To prevent tissue atropy followingtransplantation (e.g. Breast reconstructionafter mastectomy in breast cancer patient)
FAT cells Regenerative SCRe-sculpt breast
liposucction
Enhancedfat cells
Stem cells provide paracrine factors (extent cells life) & ableto differentiate to fat cells thus constant supply for cells
How Stem Cell works3. Tissue repair in degenerative diseases (clinical studies) ischemic/ hypoxic tissue release SDF-1 (stem
cell homing factor and inducer for stem cellproliferation)e.g in post myocardial infarction
Stem celldifferentiate into
vascular tissue &
Stem Cell
Harvest
Inject to the
injured area
promo e
cardiomyocyte
regeneration
How Stem Cell works4. Induction of Immune tolerance Mesenchymal Stem Cell in autoimmune,
Prevention and treatment of GVHD(in clinical studies)
Inflammatory bowel disease
5. Organ regeneration fortransplantation e.g. Regeneration of insulin producing
-pancreas, cornea, etc (Still in pre-clinical research)
,Rheumatoid Arthritis
Multiple Sclerosis
Autologous transplantation
Patient receive his/her own stem cells =Autologous
Allogeneic transplantation
PBMC/
BM/ UCB
Patient receive his/her stem cells from a donor =Allogeneic
Example for Clinical Application hematological malignancy (Leukemia)
abnormal blood cells 1st wiped out bychemotherapythen repopulated by the new stock by iv infusion
of hematopoetic stem cells n use ce s requre or a equa e engra men
>5 x 106 CD34+cells/kg body weight
7/28/2019 CTS StemCell Lecture
6/8
6
Example for Clinical Application hematological malignancy (Leukemia)
abnormal blood cells 1
st
wiped out bychemotherapythen repopulated by the new stock by iv infusion
of hematopoetic stem cells n use ce s requre or a equa e engra men
>5 x 106 CD34+cells/kg body weight
donated cells are preferably froma different donor with matchedHLA (HLA-A, -B, and DR)
Basic Immunology for Stem transplantation
What is HLA
(human leukocyte antigen) ?
Basic Immunology for Stem transplantation
HLA / human leukocyte antigen
1. Proteins present on the cell surface2. An Identity recognized by our immune system
HLA
T cell
Basic Immunology for Stem transplantation
What is the difference in stem
cell transplantation vs blood
Basic Immunology for Stem transplantation
1. In Stem Cell transplantation, the cellswill live indefinitelywithin our body
What is the difference in stem cell
transplantation vs blood transfusion ?
ong encounters wt ost ce s
2. In Blood transfusionmajority willeventually perished(onlyshort period in circulation)
Human Leukocyte Antigen
Class I Class II
HLA-A & HLA-B HLA-DR & -DQ
StimulateC totoxic T cells
StimulateT hel ercells
Incompatibility resultsin Cell Destruction
Induction ofAntibody
Production
non-matched HLA type between donor and therecipient will result in cell damage called cell rejection
7/28/2019 CTS StemCell Lecture
7/8
7
Human Leukocyte Ant igen
HLA-Atypes
e.g. -A*01, -A*34
HLA-Btypes
e.g. -B*22, -B*40
Al l humans inher it 1/2 of their enti re genetic
make-up, thus of their HLA-type, 1/2 from their
mother and the other 1/2 from their father
Human Leukocyte Antigen
Class I HLA-A
types
matchedDonor -A*01, -A*34
- * - *- , -
The degree of HLA compatibility between
donor and recipient will i nfluence the outcome
of the transplant.
Preferably > 6 allele (out of 8) matched
Human Leukocyte Ant igen
Al leles
1 2 3 4 5 6 7 8
1 allele mis-matched
A*22 A*34 A*20 A*22 DR*03 DR*04 DQ*03 DQ*04
7 allele matched
Human Leukocyte Antigen
Al leles
1 2 3 4 5 6 7 8
1 allele mis-matched
A*22 A*34 A*20 A*22 DR*03 DR*04 DQ*03 DQ*04
7 allele matchedOK for
transplantation
Human Leukocyte Ant igen
Al leles
1 2 3 4 5 6 7 8
* * * * * * * *
A*22 A*34 A*20 A*22 DR*03 DR*04 DQ*03 DQ*04
HLA typing is the most important way todecide if a donor's stem cells will match the
recipient
Basic Knowledge in Stem Cell banking for
clinical use
Why cryopreserve?1.For long term
storage
2.Increase Chance fo r
finding matched
donors
3.Especially for
Umbilical Cord
Blood Stem Cells otherwise discarded
7/28/2019 CTS StemCell Lecture
8/8
8
What is cryopreservation ?Our Cell contained >80% of water
Frozen water expands Formation of ice crystal causes cell damage
To prevent ice crystaldecrease the amountofwater absorbed by icecrystal
Dimethylsulfoxide/ DMSOprotect against thispenetrates the cells thenreplaces H2O (5-10%DMSO)
Next Lecture (3 x 50 minutes)
Lecture 2: Advances in Stem Cell Research
Duration 3 x 50 minutes
Weekly objectives:
1. Alternative sources for stem cell isolation2. Embryonic stem cell (In vitro Fertilization,
Somatic Cell Nuclear Transfer, Parthenogenesis)3. Induced pluripotentstem cell (iPS)4. Stem cell application in clinical trials