STEM SEL DAN PRODUK STEM

SEL UNTUK PENGOBATAN

REGENERATIVE
Drh. Yuda Heru Fibrianto, MP. PhD.
Bagian Fisiologi FKH-UGM Yogyakarta
Klinik Bayi Tabung “GladioolIVF” di RSIA Gladiool Magelang
PT. Dermama Indonesia
PT. Wersut Seguni Indonesia
The Stem Cell
Concept

A stem cell is an undifferentiated,

dividing cell that gives rise to a
daughter cell like itself and a
daughter cell that becomes a
specialized cell type.
Properties of a stem cell

 Must have telomerase

 Has active genes that inhibit differentiation
 Has active genes that promote growth and
cell division
 Often polarized enabling it to give rise to
disperate progeny
Embryonic Stem cell Adult Stem cell
Pluripotency

 Defined as the ability of a cell to differentiate

into any cell type of the three germ layers.
 The acquisition of pluripotency in somatic
cells requires the activation of a
transcriptional regulatory network (this
involves NANOG, SOX2, OCT4, TCF3 and
possibly others),
 Teratoma formation is the most stringent test
for pluripotency that is available in the
human system.
Embryonic Stem Cells
 Embryonic stem cells are derived from the inner cell mass
of a preimplantation blastocyst.
 These pluripotent stem cells can give rise to all tissues,
including the complete spectrum of mesoderm, endoderm,
and ectoderm derivatives
 Human ESCs were first isolated a decade ago, and have
now been demonstrated to give rise to various cell types,
including hematopoietic cells, neuron-like cells, glial
progenitors, dendritic cells, hepatocytes, pancreatic islet-
like cells, osteocytes, chondrocytes, adipocytes,
cardiomyocytes, as well as muscular, endothelial, skin,
lung, and retinal tissues.
 This quintessential differentiation potential provides a
promising avenue to produce a large quantity of
transplantable cells from a renewable source.
 Perinatal Stem Cells
 There is growing evidence to support the diverse differentiation capacity of
stem cells derived from perinatal sources, including umbilical cord blood
(UCB).
 Transplantation of UCB has been clinically successful for hematopoietic stem
cell applications resulting in high degree of engraftment, favorable
immunotolerance, and limited evidence for graft-versus-host disease
compared to adult bone marrow stem cell transplantation
 UCB-derived stem cells are capable of in vitro expansion, long-term
maintenance, and differentiation into representative cells of all three
embryonic germinal layers, that is, the endoderm (e.g., hepatopancreatic
precursor cells, mature hepatocytes, and type II alveolar pneumocytes),
mesoderm (e.g., adipocytes, chondrocytes, osteoblasts, myocytes, and
endothelial cells), and ectoderm (e.g., neurons, astrocytes, and
oligodendrocytes). (Van de Ven et al., 2007)
 UCB may provide a clinically applicable that avoids ethical challenges raised
with embryonic sources.
 Amniotic epithelial cells (AECs) from amniotic membranes can also be
induced to differentiate into diverse and specialized cell types from all three
germ layers including pancreatic cells (endoderm), cardiomyocytes
(mesoderm), and keratinocytes (ectoderm).
Adult Stem Cells

 Adult stem cells comprise a wide range of

progenitors derived from nonembryonic, non-
fetal tissues such as bone marrow, adipose
tissue, and resident stem cell pools.
 Adult stem cells are a leading candidate for
clinical application in regenerative medicine
based on accessibility, autologous status, and
favorable proliferative potential.
 Bone marrow-derived stem cells are a
cornerstone of contemporary regenerative
medicine applications
 Bone marrow-derived hematopoietic stem cells represent the
earliest example of cell-based regenerative medicine, pioneered
to address the needs of patients treated with total body
irradiation for leukemia that developed life-threatening
infections and irreversible tissue destruction.
 Stem cells defined by expression of the CD34 surface marker can
also be obtained via peripheral blood leukapheresis for clinical
engraftment.
 Provided the foundation for autologous and allogeneic stem cell
transplantation, and offer novel treatments for patients with
cancer, autoimmune diseases, and genetic diseases, including
severe combined immunodeficiency and thalassemia.
 Transplant studies have also revealed engraftment of
nonhematopoietic cell lineages derived from donor bone
marrow, unmasking subpopulations capable of a diverse range of
lineage-specific differentiation.
 Mesenchymal stem cells
 Also discovered in the bone marrow, although at low frequency compared to the
hematopoietic pool, isolated from connective components of various postnatal
tissues including adipose and synovial tissue, as well as from peripheral and cord
blood.
 Mesenchymal stem cells exhibit properties of multipotency, with the capacity to
contribute to regeneration of bone, cartilage and muscle, tissues of mesodermal
origin (Phinney and Prockop , 2007)
 Evidence also supports the contribution of mesenchymal stem cells to liver and
pancreatic islet cell regeneration, and protection in the setting of kidney, heart,
or lung injury.
 Mesenchymal stem cells secrete a spectrum of bioactive molecules that provide
a regenerative microenvironment to limit the area of damage and to mount a
self-regulated regenerative response.
 Autologous and allogeneic mesenchymal stem cells have been tested in recent
clinical trials including for treatment of osteogenesis imperfecta, Crohn’s
disease, and graft-versus-host disease.
 Bioengineered Stem Cells
 Embryogenesis is a sequential process of differential gene expression
dictated by the epigenetic environment. Exploiting epigenetic
influence on phenotypic outcome, biotechnology platforms are
developed for reversal of differentiation to achieve genetic
reprogramming of adult sources back to an embryonic state.
 Such platforms include “therapeutic cloning” and “nuclear
reprogramming” that bypass the need for embryo extraction to
generate pluripotent stem cell phenotypes from autologous sources.
 Reprogramming of adult stem cells to generate customized
embryonic-like stem cells offers the future for patient-specific
regenerative therapies
 The following three issues are important purposes of ES and iPS cell
researches for regenerative medicine: 1) Dissection of differentiation
mechanisms, 2) Application to cell transplantation, and 3) Drug
discovery
 Embryo-like stem cell
 Embryonic stem cell from somatic cell
nuclear transfer (SCNT): therapeutic
cloning
 Partenogenesis
 iPS (induced pluripotent stem cell)
 1

Collection of G0 stage

cell donor Culture Nuclear

Transfer 5
Patient 3 4
2 _ +

Enucleation
Oocyte Donor In vitro
Maturation Electrofusion

Chemical 6
activation
In vitro Culture
Embryo development

7 8
9
Imunological
5-6 hari
compatible

Embryonic stem cell

Patient

Cloned human ES cells for therapeutic process

iPS production with Transcription
Factor Transfection
OCT 3/4 C-Myc Sox 2 Nanog

Retrovirus vector
AP-staining

62 Day
Alasan pemilihan sel mesenchimal dan sel
lemak sebagai sumber sel punca

 Sifat autologous (Bang et al., 2005; Jung et al.,

2009; Kang et al., 2006).
 MSC mudah diperoleh dari jaringan pasien
sendiri, termasuk sumsum tulang, jaringan
adiposa, tulang rawan, sinovium, periosteum,
otot, dan amandel palatine dan dapat diperluas
sampai berdiferensiasi menjadi garis keturunan
sel tertentu.
 Cont...
 Sejumlah penelitian telah menunjukkan bahwa sumsum
tulang berisi populasi sel punca mesenchymal yang mampu
berdifferensiasi menjadi beberapa garis keturunan,
termasuk jaringan neuron dan glia (Jiang et al., 2002; 2003;
Nagai et al., 2007; Kim et al., 2008).

 Pengadaan sumsum tulang sangat menyakitkan bagi

pasien, jumlah sel yang dipanen hasilnya rendah dan
terbatas oleh usia pasien.

 Sel punca mesenchimal asal jaringan adiposa adalah

sumber alternatif yang baik untuk terapi sel karena akses
mudah dan jumlah sel yang tinggi serta mempunyai respon
yang baik pada kultur untuk pemulihan kemampuan
proliferasi dan memperbaharui diri sendiri dibanding
dengan MSC dari sunsum tulang (Efimenko et al., 2011).
 Cont ...
 Jaringan lemak, seperti juga sumsum tulang, berasal
dari mesoderm embrionik dan mengandung populasi
sel stroma heterogen yang mencakup adipocytes
masak, preadipocytes, fibroblas, pembuluh darah sel-
sel otot polos, sel endotel, monosit / makrofag, dan
limfosit (Caspar-Bauguil et al., 2005).
 Sel punca yang diturunkan dari jaringan lemak
(ADSCs) mempunyai kemampuan untuk
memperbaharui diri sendiri dan dapat berdifferensiasi
sepanjang garis keturunan beberapa jaringan
mesenkim, termasuk adipocytes, osteoblast, miosit,
kondrosit, sel endotel, dan kardiomiosit (Peng et al.,
2008; Lin et al., 2008; Park et al., 2008; Zuk et al.,
2002).
 Dapat berdiferensiasi menjadi sel saraf fungsional
menggunakan bFGF dan forskolin (Jang et al., 2010).
 Differentiation of HESC in vitro
Growth factors Non-protein Feeder layer
factors
Neurons NGF, EGF, bFGF, PDGF, IGF- RA MS5 (stromal cell) feeder
1, NT-3, BDNF, TGFα, fgf-8, MedII (human hepato carcinoma) condition
SHH media
Glia EGF, BFGF, PDGF, IGF-1, RA
NT-3, BDNF

Kidney NGF

Bone Dexamethasone,
ascorbic acid, β-
glyserophosphate
Cardiomyocyte TGFβ 5-aza-2’-deoxycytidine END-2 (visceral endoderm) feeder layer

Endothelial cell VEGF, PDGF

Blood cells Cytokine + BMP4 + VEGF C166 (yolk sac endothelia)

S17 (bone marrow) feeder layer

Liver NGF, HGF. aFGF Sod. butyrat Primary hepatocyte condition media

Pancreas NGF Various hormone, growth factor and matrix component

NGF: nerve growth factor; EGF: epidermal; aFGF: acidic fibroblast; bFGF: basic fibroblast; FGF-8: fibroblast;
PDGF: platelet derived; HGF: hepatocyte; IGF-1: insulin like; NT-3: neurotropin 3; BDNF: brain derived;
TGFα: transforming; TGFβ; RA: retinoic acid; VEGF: vascular endothelial; BMP: bone morphogenic protein
Stem cell homing
Schematic representation of
some of the interactions
between HSC and their
endosteal and vascular
niches.

Schematic representation of some of the interactions between HSC and

their endosteal and vascular niches.
Mechanisms of stem cell function
after homing into the damaged heart
Application and research of
stem cells
 Embrionic stem cell (ESC) yang menurunkan progenitor sel
saraf (ESNPs) adalah satu di antara kandidat jenis sel saraf
yang paling menjanjikan yang sedang diselidiki untuk
memperbaiki SSP
 Sel mampu mempertahankan potensi untuk berkembang
biak dan berdiferensiasi menjadi subtipe neuron dan glia
setelah pencangkokan (Tabal et al., 2005), dengan hasil
tertentu tergantung pada arahan lingkungan lokal
(Carpentino et al., 2008; Maisano et al., 2009).
 Begitu sel berdiferensiasi, akan membentuk neuron
fungsional yang mampu berhubungan dan menyatu ke
dalam otak penerima (Ruschenschmidt et al., 2005).
 PLoS One. 2011 Mar 4;6(3):e17560
Functional integration of grafted neural stem cell-derived
dopaminergic neurons monitored by optogenetics in an in
vitro Parkinson model.
Tønnesen J, Parish CL, Sørensen AT, Andersson A, Lundberg C, Deisseroth K,
Arenas E, Lindvall O, Kokaia M.
Neurosurgery. 2011 Jan;68(1):213-22; discussion 222.
Predifferentiated brain-derived adult human progenitor cells migrate
toward ischemia after transplantation to the adult rat brain.
Olstorn H, Varghese M, Murrell W, Moe MC, Langmoen IA.
The adult human brain contains neural stem/progenitor cells (AHNPCs) that can survive
transplantation into the adult rat brain, migrate toward a lesion, and display limited
neuronal differentiation in vivo

Hartman et al. (2010) menggunakan model tikus dari epilepsi lobus temporal,
telah teramati bahwa progenitor sel saraf yang diturunkan dari sel embrionik
(ESNPs) bertahan dan berdifferensiasi dalam lapisan sel granula setelah
melepaskan stereotaxic ke lekukan dentate (DG), menggantikan sel-sel
endogen blade atas. ESNPs dicangkokkan ke DG menunjukkan migrasi luas
melalui blade atas, sepanjang sumbu septotemporal dari hippocampus
 Epilepsia. 2010 Jul;51 Suppl 3:71-5.
 Effect of neuronal precursor cells derived from
medial ganglionic eminence in an acute
epileptic seizure model.
 Calcagnotto ME, Ruiz LP, Blanco MM, Santos-Junior JG, Valente MF, Patti C,
Frussa-Filho R, Santiago MF, Zipancic I, Alvarez-Dolado M, Mello LE, Longo
BM.

Epilepsia. 2010 Jul;51 Suppl 3:66-70.

Grafting of GABAergic precursors rescues
deficits in hippocampal inhibition.
Calcagnotto ME, Zipancic I, Piquer-Gil M, Mello LE, Alvarez-Dolado M.

Stem Cells. 2010 Jul;28(7):1153-64.

Medial ganglionic eminence-derived neural stem cell grafts ease
spontaneous seizures and restore GDNF expression in a rat model of
chronic temporal lobe epilepsy.
Waldau B, Hattiangady B, Kuruba R, Shetty AK.
 ScienceDaily (Aug. 30, 2008) — Oregon Health & Science
University scientists have successfully produced functional
auditory hair cells in the cochlea of the mouse inner ear.
The breakthrough suggests that a new therapy may be
developed in the future to successfully treat hearing loss.
The results of this research was recently published by the
journal Nature.
ScienceDaily (Aug. 3, 2009) — University of Florida
researchers were able to program bone marrow stem
cells to repair damaged retinas in mice, suggesting a
potential treatment for one of the most common causes
of vision loss in older people.

Human embryonic stem cells modified to glow red when the

stem cells become red blood cells. ScienceDaily (Nov. 29,
2009) — Victorian stem cell scientists from Monash
University have modified a human embryonic stem cell
(hESC) line to glow red when the stem cells become red
blood cells.
 Modification of radiation-induced oral mucositis (mouse) by
adult stem cell therapy: single-dose irradiation. Schmidt
M1, Piro-Hussong A, Siegemund A, Gabriel P, Dörr W.in Radiat
Environ Biophys. 2014 Jun 15.

 Mol Cancer Ther. 2014 Jun 13. pii: molcanther.0175.2014.

Human umbilical cord blood- derived mesenchymal stem
cells producing IL-15 eradicate established pancreatic tumor
in syngeneic mice. Jing W1, Chen Y2, Lu L3, Hu X4, Shao
C4, Zhang Y4, Zhou X4, Zhou Y4, Wu L5, Liu R4, Fan K6, Jin G4.

 Cardiol Rev. 2014 Jun 12.

 Cardio-Oncology Issues among Pediatric Cancer and Stem
Cell Transplant Survivors. Hochberg JC1, Cairo MS, Friedman
DM.
 Int J Hematol. 2014 Jun 14.
 Combination of bortezomib, thalidomide, and
dexamethasone (VTD) as a consolidation therapy after
autologousstem cell transplantation for symptomatic multiple
myeloma in Japanese patients. Takashima S1, Miyamoto
T, Kadowaki M, Ito Y, Aoki T, Takase K, Shima T,Yoshimoto G, Kato
K, Muta T, Shiratsuchi M, Takenaka K, Iwasaki H, Teshima T,Kamimura
T, Akashi K.

 Blood Cells Mol Dis. 2014 Jun 9. pii: S1079-9796(14)00054-0. doi:

10.1016/j.bcmd.2014.05.003. Hematopoietic stem cells and
liver regeneration: Differentially acting
hematopoietic stem cell mobilization agents reverse
induced chronic liver injury.
 Tsolaki E1, Athanasiou E2, Gounari E1, Zogas N1, Siotou E2,Yiangou
M3, Anagnostopoulos A2,Yannaki E4.
HASIL REPRODUKSI
Tekhnik kedokteran tinggi
Sekretome
 Faktor-faktor yang diproduksi oleh sel stem tersebut disebut
sekretome, mikrovesikel, atau eksosome.
 Bahan-bahan yang digunakan untuk bertahan hidup dan tumbuh.
 Beberapa penelitian terhadap faktor-faktor yang disekresikan
oleh sel stem memperlihatkan, telah mampu memperbaiki
jaringan/organ yang semula dalam kondisi rusak.
 Fakta bahwa sel stem mensekresikan bermacam-macam faktor
pertumbuhan juga telah diperlihatkan oleh para peneliti
proteomik, dengan membuktikan keberadaan berbagai macam
faktor pertumbuhan dan sitokin lainnya dalam ekstrak media
penumbuh sel stem
 Platelet Derived Growth Factor (PDGF), Interleukine (IL) – 6 dan
15, Transforming Growth Factor (TGF) β, Epidermal Growth Factor
(EGF), dan Leukimia Inhibitory Factor (LIF)
 After 1 months

After 2 months

After 44 months
After months
After 3 months
 Acne Papulo
Pustular + Scar

Courtesy by dr Titi Moertolo

 1 week post
Dermaroller
Acne Scar & Box scar

2 weeks after Skin Needling + MShCs

 Chronical Ulcer
diabeticum > 3 years

1 week

1 month after Mesenchymal product

(Injections and Cream) – Men 74 years
old
Leprosy after 30 years +
3 months burning ulcer 3 weeks after MSCs
+ Infection Product Injections +
Cream

BY Indah Yulianto
Luka bakar + infeksi 5 bulan y.l. intra kutan produk MShCs

regenerasi

Hari ke 15 : penyembuhan 75% tanpa Cicatriks

Pengolesan Krem + Produk MShCs ataupun Retraksi persendian
Fistula supra ani
Duchene
muscular
dysthrophy
syndrome
 Car
accident

After 3 times
 Diabetes Melitus
 Osteoarthritis
 DMD syndrom
 Cerebral palsi