cts stemcell lecture

7/28/2019 CTS StemCell Lecture

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Week 8

STEM CELLS

Carolin e Tan Sardjono , dr., PhD

[email protected]

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.


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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


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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


fibroblast-like

structure

100m100m

Criteria to define MSC


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


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Adult Stem Cell Sources

Collected fromumbilical vein

3. Cord Blood Whycollectedfrom


Cord Blood collection

After the baby wasborn

Collection fromumbilical vein atthe placentas part

Lipoaspirate


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


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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


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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) ?


HLA / human leukocyte antigen

1. Proteins present on the cell surface2. An Identity recognized by our immune system

HLA

T cell


What is the difference in stem

cell transplantation vs blood


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


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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


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


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


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


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


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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

cts stemcell lecture

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