conditioned medium – of adipocyte derived stem cells
DESCRIPTION
Stem Cell Therapy uses the most advanced method called AAPE / MSCIt's the most advanced method there is in the world, don't miss itTRANSCRIPT
ADIPOCYTE DERIVED STEM CELLS AND Mesenchymal Product as REGENERATIVE
MEDICINE
Indah Yulianto.,Endra Yustin., (Sebelas Maret University)., Titi Moertolo (Trisakti University).,Dewi Margono (Medical Dentist Indonesia University).,Yuyun R A(National Seoul University).,Yuda H Fibrianto (Medical Veterinary Gajah Mada University)
By definition, a stem cell is characterized by its ability to self-renew and its ability to differentiate along multiple lineage pathways.
Ideally, a stem cell for regenerative medicinal applications should meet the following criteria:
Regenerative Medicine :Uses the body’s own stem cells and growth factors is an alternative therapeutic strategy for repair of damage tissue, and becoming predominant cell base therapy
Can be found in abundant quantities (millions to billions of cells)
Can be harvested by a minimally invasive procedure
Can be differentiated along multiple cell lineage pathways in a regulatable and reproducible manner.
Can be safely and effectively transplanted to either an autologous or allogeneic host
Regenerative medicinal applications should meet the following criteria:
Human subcutaneous adipose tissue depots as a potential source of adult or somatic stem cells;
With the increased incidence of obesity in the Indonesia and abroad, subcutaneous adipose tissue is abundant and readily accessible.
Approximately 400,000 liposuction surgeries are performed all over the world each year,
These procedures yield anywhere from 100 mL to >3 L of lipoaspirate tissue
Hasil lemak dari liposuction
pellet
Adipocyte after 24 hours
Adipocyte after 24 hours
ADSC pasage 1 : hari ke 3
A. ADSCs hari ke 7 dan B. ADSC hari ke 14
A B
by dr Indah Yulianto – Yuyun Rindiastuti – Dewi M
Transdifferensiasi ADSCs to Odontoblast(immunocitokimia)
Ditujukan untuk regenerasi dan renewal Pulpitis
Courtesy by Indah Yulianto – Yuyun Rindiastuti – Dewi M
Preparing product Mesenchymal from Adipocyte
ADSC which are a population of pluripotent mesenchymal cells and possess the ability to differentiate into various lineages
ADSCs secrete various growth factors that repair and replace the defective surrounding cells
Also a good raw material for skin regeneration, reepithelialization, wound healing and wrinkling care instead of ADSC itself
Advance ADSCs Protein Extract by Definitions
Oxygen levels & exogenous
oxidative stress
Genetic stability
Cytogenetic analysis, gene
expression, bioenergetics
studies
Culture grown in physiologycal 20% oxygen
Culture grown in standard 3%
oxygen
-transcriptional activation in HIF 1A
target genes- Inhibition oxygen
consumption
- Increase genetic instability
- Decrease glycolysis
Oxygen sensing,
HIF 1 response
RESULTGrowth of hMSC at 20% O2 reduces lifespan, increases oxidative stress and the rate of telomere shortening
Telomerase shortening &
cell senescence
Cell growth
Growth of hMSC at 20% O2 increases DSB generation and chromosomal instability.
DSB level (53BP1 marker)
DNA damage & chromosomal
aberration
Exogenous source of ROS cause growth
defects & genetic
instability
DISCUSSION
Culture of mammalian cells at 20%
oxygen
- Promotes DNA damage & senescence
- Chromosomal aberration during long term culture
Importance of biosafety
in cell therapy
Culture ADMSC at 3% 0xygen
Increase growth rate
- No evidence of spontaneous immortalization
-- reduce telomere shortening rate decrease structural chromosomal aberration
Significant upregulation of gene expression in cells grown at 3%
oxygen tension related to HIF 1A stabilization
Balance bioenergetic
s pattern
Implication of gene ontology analysis (HIF
1 dependent gene expression)
Autocrine & paracrine effect in
cell growth & senescence
HIF 1A decreases OXPHOS system by
downregulating mitochondrial ATP
production & oxygen consumption
Analysis Method
Cell number & viability
Trypan blue stained cell counted by automatic cell
counting machine (invitrogen)
MSC marker CD 44, CD 106 PCR
Energy metabolism gene related glycolisis
mechanism
HIF 1 A, HIF 1B, GLUT-1 PCR
Pluripotency marker
HIF 2A, Oct-4, Sox-2, Rex-1
PCR
Courtesy by Yuyun Rindiastuti
RESULT
Cell proliferation & viability
syn-chro-
nization
Normoxic 24h
Hypoxic 24h
Normoxic 48h
Hypoxic 48h
Normoxic 72h
Hypoxic 72h
0
500,000
1,000,000
1,500,000
2,000,000
2,500,000
3,000,000
3,500,000
4,000,000
4,500,000
5,000,000
Graph 1. Cell number profile
Cell
num
ber
syn-chro-
nization
Normoxic 24h
Hypoxic 24h
Normoxic 48h
Hypoxic 48h
Normoxic 72h
Hypoxic 72h
86%
88%
90%
92%
94%
96%
98%
Graph 2. Cell viability
Cell
via
bili
ty
Courtesy By Yuyun RA
MSC surface marker expression
HIF 1A & GLUT 1 expression
Normoxic group Hypoxic group
Courtesy By Yuyun RA
3 days after skin needling + MCSs + product
Acne and Acne Scar 1 week After Skin Needling + MCSc + Product
1 week After Skin Needling + MCSs + Product
Acne and Acne Scar
Courtesy by Titik Moertolo
Alopecia Universalis : 1 month after MChSc and Product
Alopecia Universalis :1 month after MCSc + Product MSCc
By Indah Yulianto
After 2 Months
After :2 Months
By Indah Yulianto
After 4 months
After 4 months
By Indah Yulianto
(1)Luka iris dan (2) penyembuhan 2 hari pasca olesan krem produk
MCSc
1
2
Courtesy by dr Titik Moertolo
Acne Papulo pustular + Scar Acne : before and 1 week after AAPE
(3) Scar acne pasca skin needling + product & (4) 1 week after skin needling + MCSs +_ Product
34
Acne Papulo Pustular + Scar
Courtesy by dr Titik Moertolo
1 month after Skin Needling + MShCs
Acne Scar & Box scar 1 week post Dermaroller
Acne Scar before (Box Scar)2 weeks after Skin Needling +
MShCs
Courtesy by dr Titik Moertolo
Infant 2 weeks, with fistula supra ani
Tissue regeneration after 1 week using MshCs Product - Cream
BY Indah Yulianto
Chronical Ucler diabeticum > 3
years
Day three, after injection with
MSCHs product
1 week1 month after Mesenchymal
product ( Injections and Cream) – Men 74 years old
By Indah Yulianto
Leprosy after 30 years + 3 months burning ulcer +
Infection
3 weeks after MSCs Product
Injections + Cream
BY Indah Yulianto
Chronical Ulcer in M Hansen (Malperforan dupied) more than 20
years
1 week after
2 weeks after (painful & sensitivity +) 25 % reduce in 2 weeks
Bone
Luka bakar + infeksi 5 bulan y.l.
intra kutan produk MShCs
Pengolesan Krem + Produk MShCsHari ke 15 : penyembuhan 75% tanpa Cicatriks ataupun Retraksi persendian
regenerasi
Suport By : Derm Ma Ma Biotechnology Laboratory