APLIKASI TERAPI

STEM CELL

AG. Soemantri

BAGIAN ILMU KESEHATAN ANAK

FAKULTAS KEDOKTERAN UNDIP / RSUP DR KARIADI
SEMARANG
 PENDAHULUAN (1)
UU RI No. 36 tahun 2009 tentang kesehatan,
yang menyangkut masalah transplantasi
Pasal 64 ayat 1, ayat 2, ayat 3
Pasal 65 ayat 1, ayat 2, ayat 3
Pasal 66
Pasal 70 ayat 1, ayat 2, ayat 3
 PENDAHULUAN (2)
1. “Frischzellenthherapeutica” (i.e. cell
xenotransplants) are outside of regulatory
controls of the State because they are neither
drugs nor (therapeutica”, (“arzneimittel” in
German), but are another form of medication,
individually prepared by a physician, for his
specific patient, by agreement with his patient,
confirmed by the signed informed consent
prescribed by law (February, 16, 2000)
2. They are prepared for one time use only
 PENDAHULUAN (3)

3. They have no “shelf-life”

4. They are not distributed through the
usual channels, i.e. through pharmacies
5. They have to be implanted on a date
set prior to the start of their preparation
6. Their implantation has to be carried out
by the same physician who wrote a
prescription for the stem cell
xenotransplants for the named patient.
 PENDAHULUAN (4)

BUNDESVERFASSUNGSGERICHT
Der Bund ist nach Art. 74 Abs. 1 Nr. 19 GG nicht
befugt, die Herstellung solcher Arzneimittel zu
regeln, die der Arzt zur Anendung bei eigenen
Patienten herstellt (February, 16, 2000)
The International Xenotransplantation
Association Consensus Statement on Conditions
for Undertaking Clinical Trials of Porcine Islet
Products in Type 1 Diabetes (July, 27, 2009)
 DEFINISI STEM CELL
(SEL PUNCA)
Stem Cell adalah sel yang mampu memperbanyak diri
dalam waktu yang lama dan dapat berkembang menjadi
berbagai sel khusus / spesifik yang membentuk jaringan.
Potency
kemampuan berdeferensiasi kearah sel tertentu
Self-renewal
kemampuan memperbanyak melalui siklus dari
pembelahan sel sambil mempertahankan kondisi
undifferentiated
 KRITERIA STEM CELL
( SEL PUNCA )
1. Belum mempunyai fungsi
2. Kemampuan untuk berproliferasi menjadi sel
yang identik dengan sel asalnya
3. Berubah menjadi sel tipe lain dengan fungsi
yang spesifik
1. GENERAL ISSUES
- Stem Cell Biology
- Differentiated parental DNA Chain causes
stem cell pattern of cell-type switching in
Schizosaccharomyces pombe
- On equivalence groups and the Notch/LIN-12
- Cell Cycle Control, Cheekpoints, and Stem
Cell Biology
- Senescence of Dividing Somatic Cells
- Repopulating Patterns of Primitive Hemato
poietic Stem Cell
2. EARLY DEVELOPMENT
- The Drosophila Ovary : An in vivo stem cell
system
- Male Germ-line stem cell
- Primordial Germ cells as stem cell
- Embryonic stem cells
- Embryonal Carcinoma cells as embryonic
stem cells
- Trophoblast stem cells
3. MESODERM
- Hematopoietic stem cell : molecular
diversification and developmental
irrelationships
- Hematopoietic stem cell : Lymphopoiesis and
the problem of commitment versus plasticity
- The hemangioblast
- Mesenchymal stem cells of human adult
bone marrow
- Fate mapping of stem cell
4. ECTODERM
- Stem cell and neurogenesis
- Epidermal stem cell

5. ENDODERM
- Liver stem cell
- Pancreatic stem cell
- Stem cells in the Epithelium of the small
intestine and colon
 BASIC CONCEPT OF
CELLULAR AND
MOLECULAR BIOLOGY
DOGMA CENTRAL DNA

RNA

Protein

Mathematical Biosciences Institute (Ohio State Univ), 2 October 2003

 3 MOLECULES OF LIFE
All life depends on three critical molecules:
DNAs
– Hold information on how cell works
RNAs
– Act to transfer short pieces of information to
different parts of cell
– Provide templates to synthesize into protein
Proteins
– Form enzymes that send signals to other cells
and regulate gene activity
– Form body’s major components (e.g. hair, skin,
etc.)
ANOTHER VIEW OF CENTRAL DOGMA

A gene is expressed in 3 steps:

1) Transcription: RNA synthesis
2) Splicing: removal of intron sequence from RNA
3) Translation: Protein synthesis
 TRANSCRIPTION
• Transcription is initiated by a complex of
transcription factors binding to the
promoter site located upstream of the gene

•An enzyme, RNA polymerase II, travels

along the gradually unzipped DNA template
and polymerizes nucleotides into an RNA.

• The sequence of nucleotides on DNA

template determines the sequence on RNA
by following the rule of base-pair
complementarity, i.e., A – U, T- A, C – G, G
– C.

• Transcription continues until entire gene

is copied to RNA.

Animation Source of diagram: http://ghs.gresham.k12.or.us/science/ps/sci/ibbio/chem/nucleic

 SPLICING

pre mRNA
Exon 1 Intron 1 Exon 2 Intron 2 Exon 3

mature mRNA
 TRANSLATION (1)
• By translation, the
nucleotide sequence on
mRNA
determines the amino acid
sequence by genetic code.
• Genetic code: three base
pairs of RNA (called a codon)
determine one amino acid
based on a fixed table.
• Translation always starts at
AUG (start codon), and ends
with any of UAA, UAG, or
UGA (stop codon)
 TRANSLATION (2)
Transfer RNAs (tRNAs): small
RNA molecules. Most of the
tRNAs function as carriers of
amino acids and participate in
protein synthesis.

For example, the tRNA with the

anticodon CGG corresponds
with the codon GCC and
attaches alanine amino acid
onto the peptide chain.

Ribosome: a complex of
protein and rRNA
Animation
Source of diagram: http://www.wiley.com/legacy/college/boyer/0470003790/structure/tRNA/
 MODERN TECHNOLOGY IN MEDICINE

Modern technology: New approaches:

Diagnostic Genome Analysis:
Early Screening Expression gene
Prognostic analysis
Follow up Micro Array
Protein analysis
Human genome: New hopes:
Gene ther. in 2010 Cancer
Individualized Ther. Vascular/Heart dis.
Stem cell Diabetes
Antisense Alzheimers
Biomedical Engineering Parkinson
HOW MUCH DNA DO WE HAVE?

humans have 2 x 23
chromosomes
EACH cells contains 6
billion bases DNA
that is 1 meter of DNA
a human being has
>100.000.000.000.000
cells
that is 100 billion km of
DNA
 CELL – CELL
COMMUNICATION
Receptors
Signal transductions
Second messengers molecules
Nuclear transcription factors
Gene activations
Protein  Respons
CELLS CAN WITHDRAW FROM THE CELL CYCLE
AND DISMANTLE THE REGULATORY MACHINERY

G0 is a quiescent state
Cdks and cyclins disappear
Some cells enter G0
temporarily and divide
infrequenty (I.e.
hepatocytes)
Other differentiated cell
types (neurons) spend their
life in G0
Cell division increases the
Cells
number of somatic (body)
cells, and consists of:
– Mitosis (division of nucleus)
– Cytokinesis (division of
cytoplasm)
Apoptosis (cell death)
decreases the number of
cells.
Both cell division and
apoptosis occur during
normal development and
growth.
“Start” checkpoint
•Controls G1/S transition
•Commits cell to another
round of cell division

G2/M checkpoint
•Checks that DNA is fully
replicated

M checkpoint
•Is DNA division
(chromosome segregation)
progressing correctly?
 CHROMOSOMES
• genomic DNA is very long
• humans, 6 x 109 bp
• ~1.8 meters total length
• total length reduced via
associates with proteins
• individual DNA molecules
condensed into
chromosomes
• humans have 22 pairs +
X/X or X/Y (ie, diploid)
upstream exon intron exon downstream

Initiation codon termination codon

5’ 3’

3’ 5’
CENTRAL DOGMA OF MOLECULAR
BIOLOGY

DNA Sequence Amino

RNA protein phenotype
(splited by Acid
genes) sequence

Adapted from http://www.bioinfbook.org/

 CENTRAL DOGMA
Gene Genome

Transcription

RNA Transcriptome

Translation

Protein Proteome
 Cellularsignal transduction

Aprocessbyw hichsignalsfromoutsidethecellaretransferredinside Regulatebyproteinphosphorylation.

thecellthroughcascadeofproteinsinteraction. Involvedinallcellular
activities(death, growth, differentiation, etc.)
 REGULATORS : PROLIFERATION,
DIFFERENTIATION, APOPTOSIS, REPAIR

Genome : Transcriptome Proteome

Genes : Protein:
– Cell cycle RNA – cyclin, CDK,CDKI
– Differentiati RNA
– GF : GM-CSF,
o FGF
– Apoptosis RNA – Bcl-2, p53,
– Repair RNA caspase
– Metabolism – Gadd, enzym
RNA
– etc repair
RNA – dll
 HOW TO UNDERSTOOD ??

GENE:
Cytogenetics Gene activ:
FISH RNA
DNA analysis Northern blot
Southern blot Gene active :
RT- PCR
+/- Protein
PCR – RNA-se protection
Assay Western Blot
specific
MLPA Micro array SDS-PAGE
Sequencing : ELISA
R-ISH
SSCP
FISH
APLIKASI STEM CELL (SEL PUNCA)

Tidak semua dibicarakan satu persatu tentang

aplikasi
Yang disinggung yang pernah dijalankan oleh
RSUP Dr Kariadi / FK UNDIP
BERDASAR ASAL SEL YANG DIPAKAI

• Autotransplantasi
Dari satu bagian tubuh ke bagian tubuh yang
lain pada individu yang sama
• Allotransplantasi
Antar individu pada spesies yang sama
• Xenotransplantasi
Antar spesies yang berbeda
 MESODERM
Sel yang berasal dari Mesoderm ini yaitu
Mesenchymal Stem Cells of Human
Adult Bone Marrow, yang digunakan untuk
cangkok sumsum tulang
- Metoda Allolog
- Metoda Autolog
Cangkok Sumsum Tulang telah dilaksanakan di
RSUP Dr. Kariadi tahun 1987, 2 kasus dengan
autolog untuk penderita dengan diagnosis AML
M(2)
Salah satu sudah lulus Sarjana Kedokteran dan
Ekonomi
Satu kasus dengan metoda allolo untuk
penderita β thallasemia homo zygote
 ENDODERM
Cangkok hati dilaksanakan di RSUP Dr Kariadi
pada tanggal 1 Oktober 2006 dengan atresia
billier, anak “U” laki-laki umur 15 bulan dengan
donor dari ibu kandungnya.
Cangkok hati II dilaksanakan di RSUP Dr
Kariadi pada Juni 2009 dengan Allagile
Syndrome, anak “A” laki-laki umur 7 tahun
dengan donor sukarela.
 XENOTRANSPLANTASI 
AMANKAH ??
Legalitas (+)
Keputusan MA German No 1 BvR420/97, European
Community Council directive 2001/83/EC, UU
seluruh negara Uni eropa
Jaminan keamanan (+)
Hewan yang dipakai (kelinci) memenuhi persyaratan
yang dikeluarkan oleh : Guidelines on Infectious
Disease Issues in Xenotransplantasi

Merupakan hewan dari galur tertutup yang diisolasi

dan diawasi dari kemungkinan tertular penyakit
(diawasi 7 turunan )
EVIDENBASED
HASIL PENELITIAN TERAPI CANGKOK STEM CELL

3000 penderita SD yang diterapi  25% bisa sekolah secara

normal
Outcome terapi : terjadi peningkatan TB, LK, IQ, indeks volume
otak, perkembangan mental, perkembangan motorik
Evaluasi mental dan psikologis 83 penderita SD yang diterapi :
terjadi penurunan hiperaktivitas, peningkatan konsentrasi dan
peningkatan kemampuan bahasa ekspresif
Evaluasi imunologis : terjadi peningkatan IgA dan penurunan IgG 
Menurunnya frekuensi dan beratnya infeksi bakteri
 EVIDENCE BASED
Pada Cangkok Stem Cell (CST) usia ≤
3½ tahun
kenaikan MDI > 50 point terjadi pada :
 17 % bila tidak di CST
 58 % pada CST I
 71 % pada CST II
Pada usia 4-9 tahun
kenaikan 25-49 point
Figure 2. Phosphorylation targets of DYRK1A. The Hsa21-encoded kinase
DYRK1A has been shown to phosphorylate a multitude of targets, which
have been implicated in a number of biological processes and DS-
associated phenotypes, including endocytosis and AD.
 RINGKASAN
Telah diutarakan konsep dasar biomolekuler,
siklus sel
Definisi dan jenis-jenis, kriteria stem cell (sel
punca)
Dasar hukum
Dasar keberhasilan asosiasi stem cell
Aplikasi yang pernah dilakukan di RSUP Dr
Kariadi/FK UNDIP : cangkok sumsum tulang,
cangkok hati, cangkok stem cell