EXOSOME
Core Technology: Reprosome
®
Reprosome
Reprogramming factors Application to cell therapy of
Ultrasound related to environmental various diseases and beauty products
characteristics in the
Dermal Exosome
Fibroblasts
Entr • Exosomes secreted from fibroblasts in the process of cellular
reprogramming by Entr® system
• Mass production of reprosome is possible
• Reprosome induction is very simple and takes a day
Various Environments • The cell penetration of reprosome is very good to regenerate tissue
Cell differentiation media
Exosome, next generation of regenerative medicine
HOT ISSUE– Possibility of regeneration only with exosomes without stem cells
- what is exosome ?
- Size: 50~200 nm
- Cell-secreted vesicles containing RNAs, miRNAs, and proteins
- In the spotlight as a next-generation regenerative medical
technology suitable for clinical application
Exosome potentials
.
Safety ↑
Anti-
inflammation Drug
delivery
Anti-
fibrosis Cellular
Regeneration penetration↑
↑
Exosomes, Very good But ----
MSC exosomes: limited application to therapy of various diseases
MSC-derived exosomes contain
RNAs and proteins mainly related
to regeneration of osteoblasts,
adipocytes, and chondrocytes.
MSC
No massive production of MSC
Adipose MSC Fibroblast
tissue 5 x 106 1 x 107
passage 11 (211=2,048) 23 (223=8,388,608)
4,096
fold Cell 10 billion (10,240 83 trillion (83,886,080 x
65,536 fold
production x 106 ) cells 106) cells
difference 823,875,840 x 106 cells
MSC has low yield of exosome
Exosome yield from 1 x 106 cells 2.0´1009 16,300 fold
1.8´1009
1.6´1009
Adipose-derived MSCs Fibroblasts treated by Entr 1.4´1009
No. of exosome particles
CD63 CD63 1.2´1009
1.0´1009
8.0´1008
6.0´1008
4.0´1008
2.0´1008
1.0´107
8.0´106
6.0´106
4.0´106
2.0´106
Reprosome A-MSC
ASCE+
(1,000 x dilution) Reprosome A-MSC
CD63 : Exosome marker (exosome particle shown in red) 평균 1.43x109 8.71x104
표준오차 ± 3.17x108 ± 6.31x103
Reprosomes address existing exosome challenges
2018 IF 15.881
A shift of thought overcomes the problems of exosome
Ultrasound-directed permeation of
Entr TM Environmental transition-guided cellular reprogramming
1. Human dermal fibroblasts: massive cell production
2. Environments: Various cell culture media for inducing
Massive amounts of
customized reprogramming factors Entr
reprosome within a day
3. Ultrasound : enables influx of environments into
fibroblasts and massive production of exosomes
Cellular composition (Environment)
Ultrasound
1 Days
HDF cells Environmental
1x106 cell/ml Permeation
10 trillion reprosomes
International peer-reviewed SCI publications of Entr technology
2018년 IF 12.479 2018년 IF 15.881 SCI IF 5.055
Biomaterials, vol.143, p.65-78 (2017) ACS NANO, vol.12, p.2531-2538 (2018) Journal of Molecular and Cellular
Cardiology, vol. 126, p.118-128 (2019)
Automation equipment for reprosome production
ü On single processing
ü Provide stable mass production of reprosome
- 2 minutes
- 10 trillion reprosome particles
ü Provide uniform production of
reprosome
Human dermal
fibroblasts (1 x 106 ) [tube for cell]
IP registered
[Entr 시스템에 의한 기능성 리프로좀 유도]
[Movie, UltraRepro equipment that incorporates [Movie, movement of reprosomes generated
Environments [Tube for media] know-how in Entr technology] using UltraRepro]
IP registered IP registered
Great skin penetration of reprosomes
Before treatment 24 hrs after reprosome treatment
Diagram of skin tissue
: Reprosome
[Movie, penetration of reprosomes through skin tissue after spraying them on skin.
*For tracking reprosomes, they were stained by Did Reprosomes are stained in red by Did]
Development of various customized reprosomes
Innovative technology
Various environments targeting
• Brain
• Liver Customized®
HDFs-based • Lung Reprosome Beauty and healthcare
• Hair follicle • Beauty products
Entr platform Induction of various types of
reprosomes containing • Hair loss
• Skin hundreds of reprogramming
• Telomere factors for each tissue
Other products
Cell therapy
Adipose-derived MSC
adipocytes, osteoblasts,
No customized • Brain
chondrocytes, muscle exosomes • Lung
• Atopy
• Liver
Reprosomes for regenerating hair follicle
Massive induction of reprosomes containing
reprogramming factors for hair follicles by Entr
Hair follicle differentiation media-based Entr Reprosomes containing various
treatment induced massive production of exosome reprogramming factors for hair follicles
< Entr treatment >
Shh/CD63 Wnt/CD63 B-catenin CD63 Krt25/CD63
Reprosome effect on regenerating hair growth and hair follicles
Great effect of hair
regeneration by reprosomes
Hair growth-promoting activity by spraying Activating hair follicle
reprosomes on skin regeneration by reprosomes
Control Reprosomes
The first success case of hair growth using a nude mouse
Reprosomes for skin regeneration
Entr inducing massive reprosomes containing skin
regeneration factors from HDFs
Embryonic stem cell media-based Entr Inducing reprosomes containing RNAs and
treatment showing production of massive proteins for skin regeneration
exosomes
Anti-oxidant and Skin regeneration
anti-aging factors factors
<. Entr treatment>
Reprosome effects on wound healing
Great effect on wound healing in C57 mice by spraying
reprosomes on skin
Reprosomes promoting the
Activation factors related to skin tissue regeneration
recovery of injured skin tissue
Activating neovascularization Activating skin cell growth
Activating RNA expression of genes related to wound healing
Reprosome effect on atopic dermatitis
Epidermal thickness
Anti-Inflammation Mechanism
Telomere
(TERT)
Only stress can retard the shortening of telomere length
Reprosome for telomere length recovery
Telomere length recovered by
Entr-based reprosomes
Reprosomes containing
telomerase after Entr Telomere length recovery and resultant anti-aging by
treatment reprosomes
39
Reprosome biodistribution in vivo
IV Injection of reprosomes stained by Did (red)
Reprosomes for liver regeneration
Reprosomes for hepatocyte induction regenerate
injured liver tissues
Hepatocyte differentiation media-based Entr induced Recovery of injured liver tissue by
reprosomes containing liver regeneration factors reprosomes
After
20 days
Hepatic differentiation media
Genes
related to
liver
regenerati
on
reprosomes for lung regeneration
Superior competitiveness of Reprosome
Reprosome Exosomes from MSC
Cell production
/donor tissue 65,536 fold 4096 fold
Therapy for 5,000 times
Adipose limited
Simple diseases:
MSC-derived extraction CD63: Exosome marker
chondrocytes
Exosome
osteoblasts,
Exosome
adipocytes production
/106 cells
10,000 times
Protein chip: 1020 proteins related to regeneration
Entr- Customized
reprosomes Therapy for Number of
inducing various
proteins for
by regeneration/
reprosomes diseases
environments 107 exomes
from HDFs 10 times
About 930 types of proteins About 80 types of protein
Clinical safety and study of Reprosome for skin regeneration
5x107 reprosomes/ml Treatment
Safety inspection by the KFDA (determined as non-toxic)
Test items Company Test result
Before
Single dose toxicity test (rat-dermal) Biotoxtech No abnormality
13-week repeated dose toxicity test (rat-dermal) Biotoxtech No abnormality
Skin sensitization test (LLNA-alternative test method) Biotoxtech No abnormality
In vitro 3T3 NRU phototoxicity test
Biotoxtech No abnormality
(alternative test method)
Photosensitization test Biotoxtech No abnormality
Return mutation test Biotoxtech No abnormality
Chromosomal abnormality test Biotoxtech No abnormality
Micronucleus test Biotoxtech No abnormality
Skin irritation test (alternative test method) Biotoxtech No abnormality
Eye irritation test (alternative test method) Biotoxtech No abnormality
After
Hypoallergenic det
Human patch test KDRI
ermination 3 Days 21 minutes 12 Days 10 Days 30 Days
COA Company Test result
Mycoplasma residual test BIOPS No abnormality
Sterility (fungal, bacterial) test BIOPS No abnormality
Virus residual test (in vitro) BIOPS No abnormality
Virus residual test (in vivo) KNOTUS No abnormality
* KDRI (Korea Dermatology Research institute)
Revenue-based system of reprosomes
Movie: cGMP production plant of reprosome ICID registration
Supplying reprosomes to hospitals
3 types of Reprosome brand
D-27, Before D-27, After
A-27, Before A-27, After
A-27, D-27, H-27,
for for for
atopy skin hair
H-27, Before H-27, After
