1. 01. Bio Master의 개요
Automated Extraction & Culture
System of Adipose-derived Stem Cells
2008.2.24-revised
Dr. H.Y.Lee, Dr.H.J.Yang, KangNam Plastic Surgery Clinic,
Medi-Khan(USA) Inc.

2. 01. Bio Master의 개요
Research & Trend of
Stem Cell

3. 01. Bio Master의 of Adult Stem Cell
Importance in Research 개요
Can Overcome Issues over Embryonic Stem Cell Research Can be Utilized as Patient-specific Stem Cell
Ethical Issue Patient-
Difficulty in Regulating Differentiation for Cell Therapy
Required Long Time before Commercial Use Over 101 millions of world-wide patients Need
world-
Cell Therapy. (Perry D., Science 287, 1423, 2000)
Superiority in Technological Competitiveness
Can Isolate Stem Cells from Fat Tissues Current Adipose-derived Stem Cell Research
Adipose-
Adult Stem Cell Stays at Early Stage. So, Easier to possess
Lots of adult stem cells exist in fat tissues Technological Superiority than other Research.
Can forecast it as Faster Growth Industry
Can Obtain Large Quantity o f Stem Cells
Expected to grow by 18.5 % to 10.9 Billion
Diversified Technologies in Isolating US Dollars by 2015.
& Culturing Adult Stem Cells (Jain PharmaBiotech Report, 2005)

4. 01. Bio Applications of개요
Potential Master의 Adipose-derived Stem Cells
The Major advantages of adipose tissues as a source of regenerative cells
• High Yield : A therapeutic dose of regenerative cells can be isolated within
about 1~2 hours without cell culture.
Safety : Patients receive their own cells (autologous-use), so there is no risk
of immune rejection or disease transmission.
Versatility : Stem cells from adipose tissues can impart benefits from
multiple mechanisms-of-action as pluripotent progenitor cells.
Spees JL, Olson SD, Ylostalo J, et al.: Differentiation, cell fusion, and nuclear fusion during ex vivo
Wound Healing repair of epithelium by human adult stem cells from bone marrow stroma.
Proc Natl Acad Sci USA.100:2397-2402, 2003
Kotaro Yoshimura, Katsujiro Sato, Noriyuki Aoi, Masakazu Kurita, Toshitsugu Hirohi and
Plastic Surgery Kiyonori Harii : Cell-assisted lipotransfer (CAL): supportive use of human adipose-derived cells for
soft tissue augmentation with lipoinjection. Aesthetic Plastic Surgery, 1432-5241, 2007
Dragoo, J.L., Samimi, B., Zhu, M., Hame, S.L., Thomas, B.J., Lieberman, J.R., Hedrick, M.H., and
Bone & Cartilage Benhaim : P.Tissue-engineered cartilage and bone using stem cells from human infrapatellar fat pads.
regeneration J. Bone Joint. Surg. Br. 85, 740, 2003.
Planat-Benard, V., Menard, C., Andre, M., Puceat, M., Perez, A., Garcia-Verdugo, J.M., Penicaud, L.,
and Casteilla : L. Spontaneous cardiomyocyte differentiation from adipose tissue stroma cells.
Neuron & Heart Circ. Res. 94, 223, 2004.
regeneration Safford, K.M., Hicok, K.C., Safford, S.D., Halvorsen, Y.D., Wilkison, W.O., Gimble, J.M., and Rice,
H.E.Neurogenic differentiation of murine and human adipose-derived stromal cells.
Biochem. Biophys. Res. Commun.294, 371, 2002.

5. 01. Bio Master의 개요
Necessity in Technological Development

6. 01. Bio in Technological Development I
Necessity Master의 개요
Increased
Autologous Fat or Stem Cell Implantation for Cosmetic or plastic Surgery
Need for
1 Autologous
Autogenous Stem cell Implantation for Burnt and Depressed Body Parts and
Nerve Regeneration
Implantation
Harvesting Fat Tissues → Isolation of Stem Cells → Massive Culturing →Implantation
to Patients (Autologous use)
Required
Massive Cell
Implantation
Isolation &
2 Culture
Collagenase
Treatment
Culturing for
Technology For 1~2 hrs Isolation after centrifugation, oil
10~20 days
removal and washing processes
It takes 2~3 hours to isolate stem cells by manual methods. Faster process is required.
The existing isolation technologies require many experts with much labor cost.
To utilize stem cells for treatment purpose, quantified data are needed.
in vivo system : Employing the optimized environment for stem cell isolation.
Required to ( Can maintain constant temperature and humidity and prevent contamination)
develop Effective Reduced time of stem cell isolation, Saved Cost, Least Labor
3 Cell Isolation With optimized technology and automated isolation system, quantified data is available.
Designed to be suitable for GMP facility or for production system of stem cells for cell
Technology therapy.

7. 01. Bio in Technological Development II
Necessity Master의 개요
Cell Banking
Increased need for organ storage for autologous
implantation.
Trial of homogenous(Heterogeneous) implantation
- Trial of cell therapy for brain, spine and heart with
low immunorejection
- Cell banking system is required for massively
extracted and stored stem cells for cell therapy.
Cell Therapy
Harvest of
highly
concentrated After massively culturing adipose-derived stem cells, differentiation of ASCs
into adypocytes, osteocytes or neurocytes for required treatment purpose.
fat tissues
Implantation into patients
Cosmetic Surgery
Isolation of Adipose-
derived Stem Cells
Transplantation of fat cells and ACSs for cosmetic surgeries like breast
augmentation, facial microinjection and wrinkle removal
Showing good surgery results from ASC implantation.

8. 01. Bio Master의 개요
Current Trends in Technological
Development

9. 01. BioTrends of Stem Cell Extractors
Current Master의 개요
In domestic market, development technology for stem cell isolation systems is at lower level.
Companies retaining technology of stem cell extraction method and system
Cytori Therapeutics, Inc.
Tissue Genesis, Inc. Osiris Therapeutics, Inc.
Macropore Biosurgery, Inc.
Biosurgery,
www.tissuegenesis.com www.osiristx.com
www.osiristx.com
www.cytorix.com/us
www.cytorix.com/us
System and method of isolation and Isolating, purifying and culturing
Retaining Method and apparatus of preparation
concentration of regenerative cells from method of human mesenchymal stem
for tissue transplantation
Technology tissues and others.
PCT No.: WO 200709036
cell( hMSC) and others.
PCT No. : WO 2005/012480 US Patent No.: US5827740
(Patent) Applied : June 25, 2004
Applied : July 12, 2006
Applied : July 30, 1996
Product CelutionTM TGI 1000 system, TGI 1001 system Condrogen, Osteocel
Consisting of collecting chamber and Applying filtering and centrifugation With various methods, trial of
processing chamber. Processing chamber simultaneously, can isolate stem cells. isolation of stem cells extracted from
bone marrow.
can apply filters to remove impurities Similar to Cytori’s CelutionTM
from fat cell mixtures during (antigen-antibody response)
centrifugation and collect stem cells.
Principle
As an isolation method of stem cells
During Centrifugation, extraction of Isolation of a certain stem cells using
Function stem cells using multi-staged filters.
using filter and centrifuge, can also
antigen-antibody response

10. 01. BioTechnological Trends of Bioreactor
Current Master의 개요
The current technologies of Bioreactors have been world-widely developed.
Commercialization of Bioreactor was made by Aastrom, an USA-based company in mid 1990.
In recent time, Bioreactors with nano scale for stem culture are under research.
Cellforcel, an automated bioreactor, was developed by Cell-force, a Japanese company (www.cell-force.com).
In Korea, reportedly Biotron Company has developed an bioreactor for stem cell culture (www.Biotron.com).

11. 01. Bio Master의 개요
Major Technologies of Medi-khan

12. 01. Bio Master의 개요
Major Technology of Medi-khan
2001 2004 2007 2010
Cell Therapy
Automated Extractor system
Of Stem Cells
Harvesting Machine
of pure fat tissues
Automated Stem Cell Extractor
Lipokit (YC-100)
Cell Therapy System
Fat Transfer Device integrating liposuction and • In 2006, development of automated
centrifugation in one system • Trial of massive culturing of stem
stem cell extractor.
Fast & Massive Fat Transfer can be performed. cells through 3 dimensional cell
• Can automatically isolate stem cells
Can harvest pure, concentrated fat tissues using culturing technology.
closed-circuit AFT syringes equipped with from fat tissues at fast speed.
• Introduction of stem cell culturing
weight-mesh piston to remove free oils and •Can separate stem cell with high yield
technology to stem cell extractor is
prevent contamination. rate at 37℃ .
KFDA Approved (2004) w/ proprietary patent under process.
•All processes are automatically made.
Sales Contract with Biomaster in Japan • With fully automated system of
• 1st Quality test was finished w/
Domestic Sale to Plastic & Cosmetic Surgeons stem cell isolation and culture, our
Domestic(15) and overseas(11) patent protected enhancing its performance.
advanced technology can be applied
•Pending Patent (#20-2004-0010685) –
for stem cell production for cell
Centrifuging unit integrating
therapy.
liposuction and fat injection. And other
patent (Domestic 9, Overseas 9)

13. 01. Bio Master의Stem Cell Extractor(YC-100)
Patents for Automated 개요
Application No. : 10-2006-0060513, Applied Date: June 30, 2006
Summary : Centrifugal device having a conical shape to move up materials on the receptor to the top layer
Centrifuging Device by centrifugal force and also having a filter on the top layer of the receptor to pass only some parts of the
materials through.
Application No. : 10-2006-0111264, Applied Date : Nov. 10, 2006
Summary: Device & Method featured by angle control part to control a receptor angle so that
Centrifuging Device & Method collected top layer fluids during centrifugation can flow down at over 90 angle.
Application No. : 10-2006-0111265, Applied Date : Nov. 10, 2006
Summary: Centrifugal device & Method to exactly and easily distinguish and recover fluid layers by
Centrifuging Device & Method difference of specific gravities and fluid resistance between the separated fluid materials
when opening part of the container is opened after revolution body is revolved by centrifugation.
Application No. : 10-2006-0111266, Applied Date : Nov. 10, 2006
Summary: Centrifugal device and method to distinguish and separate fluids by differences of large or small
Centrifuging Device & Method Centrifugal forces and resulting control of revolution speed. Based on the principle that magnitude of
centrifugal force by revolution speed differs between the specific components of fluid materials.

14. 01. Bio Master의Stem cell Extractor, YC-100
Technological Value of 개요
(International Search Report)
MEDIKAN YC-100
CYTORI THERAPEUTICS TISSUE GENESIS

15. 01. Bio Master의 개요
Automated Stem Cell Extraction
& Culture Device (YC-100)

16. 01. Bio Stem Cell Extraction & Culture Device (YC-100)
Automated Master의 개요
As the 1st stage for Automated Cell Culture System, achieved at automation of whole process of stem cell extraction.
With no separate work or additional process, can automatically separate stem cells at high speed with just one consecutive process for
experimental or therapy purpose.
Basic Specification
Basic Specification External Design of YC-100
External Design of YC-100
Items Specifications
Max RPM 3,000 rpm
Max Capacity 2,000 ml
Max centrifugal force >200g
Speed preset and 구 분 1 rpm
display
Rotor radius of rotation
0.1 rpm
range
Temperature Range -20 ℃ ~ +50 ℃
99 hrs 59 min 59 sec
Timer
And “Hold” for free run
Acceleration Time 3 stage
Deceleration Time 10 setting
Drive system Ultra-precision Servo Drive control
Programmability 100 memory
RPM, RCF, Time, Temperature, Temp,
Digital Display
Limit, Program, Acel/Decel
Power Requirements 220 V, 60 Hz, single phase, 3.0 kw
Dimension 1310 W×620 D×1230 H / mm

17. 01. Bio Stem Cell Extraction & Culture Device (YC-100)
Automated Master의 개요

18. 01. Bio Stem Cell Extraction & Culture Device (YC-100)
Automated Master의 개요
Infusion Apparatus of Fat Tissues & Reagents
• Control of infusion & discharging quantities using roller pump and pneumatic apparatus
Using simultaneously operative Upper and Lower Infusion Tubes, can infuse or discharge required fat
tissues or reagents even during operation.
Can control temperature at the mount for Fat tissue and Reagent Container
Can sense the weight of every infusion material or reagent using load-cell.
Use of disposable silicon tube

19. 01. Bio Stem Cell Extraction & Culture Device (YC-100)
Automated Master의 개요
Touch-Pad Monitor
• Window Garbin (가빈 ?) User - Graphic User Interface + Touch Screen
Key Pad : Supplementary input apparatus
With employment of touch-pad monitor, can easily and simply set up required
programs and operate YC-100.

20. 01. Bio Stem Cell Extraction & Culture Device (YC-100)
Automated Master의 개요
Built-in Revolutionary Chamber
• Chamber with non-uniform plane against centrifugal forces
• Can isolate large quantity of stem cells at a time unlike the existing extractors.
• Using difference of centrifugal force and gravity at each point, can collect stem cells at end of cone.
• With employment of double-step tilting function to the cones, can isolate stem cells very well
• Using disposable membrane within chamber, can prevent any possible contamination during isolation.

21. 01. Bio Stem Cell Extraction & Culture Device (YC-100)
Automated Master의 개요
Air Handling Unit (Heating)
Can control even minute temperatures using air conditioner and hot air blower
Can remove contaminated materials with circulatory filtering of inner air flow.

22. 01. Bio Master의 개요 & Culture System, (YC-100)
Automated Stem Cell Extraction
Stroboscope Light
Moving Image Camera
Strobo & Moving Image Camera
Synchronization of intermittent on/off timing on revolution of revolving chamber
Can observe the end part of the cones of the chamber with naked eye even during high-speed revolution.
Can shoot both stopped and moving images of rotor (?) using high-tech moving image camera
Can observe the whole process of stem cell isolation

23. 01. Bio Master의 개요 & Culture System, (YC-100)
Automated Stem Cell Extraction
Other Functions of YC-100
Based on embedded CPU, employing high-end PC
Graphic-based Interface for user-friendliness
Earth Leakage Circuit Breaking Function
Proprietary Patented Automation System

24. 01. Bio Master의 Cell extraction & Culture
Efficiency Test for Stem 개요
System, YC-100
vs.
Harvesting fat tissues by Lipokit (made by Manual Method: manual isolation of stem cells from
Medikhan Corp.) fat tissues directly by skilled expert
To minimize errors, efficiency test with only Automated Method: isolation of stem cell by YC-100
highly concentrated fat tissues having over 90%
purity to be harvested by Lipokit
Comparison & analysis of efficiency rates of stem
cells isolated by experts and by YC-100
Collagenase treatment
Analysis of over 3 times per session using same fat
Separation of fat tissues by centrifuge
tissues
Washing by PBS buffer
Testing totally over 20 sessions of efficiency rates
Cell harvest
Counting the number of total cells (ADSCs &
fibrobast etc.) isolated from fat tissues by automatic cell Cell counting
counter after staining with PI (Propidium Iodide)

25. 01. Bio Master의Cell Isolation by YC-100
Efficiency Test of Stem 개요
Manual Automated
Cell number
(0.1x million 9.65 10.9 140
cells/fat1cc)
120
Yield (%) 100 112.84
Isolation yield (%)
100
80
manual Automated
60
40
20
0
수동 자동
Isolation method
Efficiency Rate for Stem Cell Isolation by YC-100 will be continuously tested and enhanced.
Characterization (like FACS analysis, proliferation and differentiation) of isolated adipose-derived stem
cells will also be identified.

26. 01. Bio Master의 of Adipose-derived stem cell (a few processes are
Processing for Isolation 개요
intentionally omitted. Confidential.)

27. 01. Bio Master의 개요
Potential Prospect of YC-100

28. 01. Bio Master의 Technology
Development of Culture 개요
Development of 3 Dimensional scaffolds for cell proliferation
Development of scaffolds with various sizes and shapes
Development of Scaffolds for Injection
Based on PLLA scaffolds, mixed scaffolds with various biological synthetic
materials are under development.
Together with stem cell extraction & culture system, efforts for high
efficiency of stem cell culture and proliferation will be continued.
< In vitro proliferation test using ADSC P0 >
Retaining Patents (Domestic 13, Overseas 6)
10-2002-0001131 : As an injection material, solid chitosan powder with sponge structure to be
injected for human body and its production method and injection apparatus.
10-2002-0025375 : As an injection material, solid chitosan powder with sponge structure to be
injected for human body and its production method
10-2002-0064189 : In Vitro Culture Method of human cells.
10-2002-0085331 : 3 dimensional cell culture method using solid powder
x200
50
10-2005-0078382 : As an injection material, solid chitosan powder with sponge structure to be
PLLA beads
injected for human body and its production method
Cell number (1X 105 cells/beads 1cc)
45
Fat scaffold
40 Dextran beads
35 10-2006-0079725 : Micro Scaffolds with non-spherical 3 dimensional stereo structure to be
30
25
produced by rapid prototyper and used for cell culture and transfer.
20
15 10-2007-0041058 : Cell culturing scaffold with increased specific gravity and its production
10 method.
5
0
initial 6h 3d 6d 10d
Culture time

29. 01. Bio Master의 개요
Stem Cell Culturing System
<Simulation for 3 Dimensional Stem Cell Culture >

30. 01. Bio Prospect of의 개요 Cell Extraction and Culture Device
Potential Master YC-100 Stem
(Extraction of Stem Cell + Bioreactor + Cell therapy)
Exploitation of Cell Therapy
Cell Banking Business
Medikhan
Biomaster (Japan)
Development of automated
Clinical Research using Stem Cells
Stem Cell Extraction &
Culture Device
Dreaming of Future High-tech
Bio Business

31. 01. Bio Master의 개요
FDA regulation on HCT/P’s

32. 01. Bio Master의 개요
FDA regulation on HCT/P’s

33. 01. Bio Master의 개요
THANK YOU