01. 01. Bio MasterBio Master의의 개요개요
Automated Extraction & Culture Automated Extraction & Culture System of AdiposeSystem of Adipose--derived Stem Cellsderived Stem Cells
2008.2.242008.2.24--revised revised
Dr. H.Y.Lee, Dr.H.J.Yang, Dr. H.Y.Lee, Dr.H.J.Yang, KangNamKangNam Plastic Surgery Clinic, Plastic Surgery Clinic, MediMedi--Khan(USA) Inc.Khan(USA) Inc.
01. 01. Bio MasterBio Master의의 개요개요Importance in Research of Adult Stem Cell
Can forecast it as Faster Growth IndustryCan forecast it as Faster Growth IndustryExpected to grow by 18.5 % to 10.9 Billion Expected to grow by 18.5 % to 10.9 Billion US Dollars by 2015.US Dollars by 2015.(Jain (Jain PharmaBiotechPharmaBiotech Report, 2005)Report, 2005)
Can be Utilized as PatientCan be Utilized as Patient--specific Stem Cell specific Stem Cell for Cell Therapyfor Cell Therapy
Over 101 millions ofOver 101 millions of worldworld--wide patients Needwide patients NeedCell Therapy. (Perry D., Science 287, 1423, 2000)Cell Therapy. (Perry D., Science 287, 1423, 2000)
Superiority in Technological CompetitivenessSuperiority in Technological Competitiveness
Current AdiposeCurrent Adipose--derived Stem Cell Researchderived Stem Cell ResearchStays at Early Stage. So, Easier to possess Stays at Early Stage. So, Easier to possess Technological Superiority than other Research.Technological Superiority than other Research.
Can Obtain Large Quantity o f Stem CellsCan Obtain Large Quantity o f Stem Cells
Diversified Technologies in IsolatingDiversified Technologies in Isolating& Culturing Adult Stem Cells& Culturing Adult Stem Cells
Can Overcome Issues over Embryonic Stem Cell ResearchCan Overcome Issues over Embryonic Stem Cell ResearchEthical Issue Ethical Issue
Difficulty in Regulating DifferentiationDifficulty in Regulating DifferentiationRequired Long Time before Commercial UseRequired Long Time before Commercial Use
Can Isolate Stem Cells from Fat Tissues Can Isolate Stem Cells from Fat Tissues
Lots of adult stem cells exist in fat tissuesLots of adult stem cells exist in fat tissuesAdult Stem Cell
01. 01. Bio MasterBio Master의의 개요개요Potential Applications of Adipose-derived Stem Cells
Wound Healing
Plastic Surgery
Bone & Cartilage regeneration
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 riskof 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 repair of epithelium by human adult stem cells from bone marrow stroma. Proc Natl Acad Sci USA.100:2397-2402, 2003
Neuron & Heart 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.
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. Circ. Res. 94, 223, 2004.
Kotaro Yoshimura, Katsujiro Sato, Noriyuki Aoi, Masakazu Kurita, Toshitsugu Hirohi and 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 Benhaim : P.Tissue-engineered cartilage and bone using stem cells from human infrapatellar fat pads.J. Bone Joint. Surg. Br. 85, 740, 2003.
01. 01. Bio MasterBio Master의의 개요개요Necessity in Technological Development I
Required Massive Cell Isolation &
Culture Technology
Required Massive Cell Isolation &
Culture Technology
Required to develop Effective
Cell Isolation Technology
Required to develop Effective
Cell Isolation Technology
1
2
3
Increased Need for
AutologousImplantation
Increased Need for
AutologousImplantation
Autologous Fat or Stem Cell Implantation for Cosmetic or plastic SurgeryAutogenous Stem cell Implantation for Burnt and Depressed Body Parts and Nerve Regeneration
Harvesting Fat Tissues → Isolation of Stem Cells → Massive Culturing →Implantationto Patients (Autologous use)
in vivo system : Employing the optimized environment for stem cell isolation. ( Can maintain constant temperature and humidity and prevent contamination)
Reduced time of stem cell isolation, Saved Cost, Least LaborWith 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 celltherapy.
CollagenaseTreatment
For 1~2 hrs Isolation after centrifugation, oil removal and washing processes
Culturing for 10~20 days
Implantation
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.
01. 01. Bio MasterBio Master의의 개요개요Necessity in Technological Development II
Harvest of highly
concentrated fat tissues
Isolation of Adipose-derived Stem Cells
Cell BankingIncreased 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
After massively culturing adipose-derived stem cells, differentiation of ASCsinto adypocytes, osteocytes or neurocytes for required treatment purpose.
Implantation into patients
Cosmetic Surgery
Transplantation of fat cells and ACSs for cosmetic surgeries like breast augmentation, facial microinjection and wrinkle removal
Showing good surgery results from ASC implantation.
01. 01. Bio MasterBio Master의의 개요개요Current Trends of Stem Cell Extractors
Companies retaining technology of stem cell extraction method anCompanies retaining technology of stem cell extraction method and systemd system
CytoriCytori Therapeutics, Inc.Therapeutics, Inc.MacroporeMacropore BiosurgeryBiosurgery, Inc., Inc.
www.www.cytorixcytorix.com/us.com/us
Tissue Genesis, Inc.Tissue Genesis, Inc.
www.tissuegenesis.comOsiris Therapeutics, Inc.Osiris Therapeutics, Inc.
www.www.osiristxosiristx.com.com
Retaining Retaining TechnologyTechnology
(Patent)(Patent)
System and method of isolation and concentration of regenerative cells from
tissues and others. PCT No. : WO 2005/012480
Applied : June 25, 2004
Method and apparatus of preparation for tissue transplantationPCT No.: WO 200709036
Applied : July 12, 2006
Isolating, purifying and culturing method of human mesenchymal stem
cell( hMSC) and others.US Patent No.: US5827740
Applied : July 30, 1996
Condrogen, Osteocel
With various methods, trial of isolation of stem cells extracted from
bone marrow. (antigen-antibody response)
Isolation of a certain stem cells using antigen-antibody response
ProductProduct CelutionTM TGI 1000 system, TGI 1001 system
PrinciplePrinciple
Consisting of collecting chamber and processing chamber. Processing chamber
can apply filters to remove impurities from fat cell mixtures during
centrifugation and collect stem cells.
Applying filtering and centrifugation simultaneously, can isolate stem cells.
Similar to Cytori’s CelutionTM
FunctionFunction During Centrifugation, extraction of stem cells using multi-staged filters.
As an isolation method of stem cells using filter and centrifuge, can also
perform even implantation at a time
In domestic market, development technology for stem cell isolation systems is at lower level.
01. 01. Bio MasterBio Master의의 개요개요Current Technological Trends of Bioreactor
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).
01. 01. Bio MasterBio Master의의 개요개요Major Technology of Medi-khan
Lipokit
Cell TherapysystemCell TherapysystemAutomated Extractor
Of Stem CellsAutomated Extractor
Of Stem CellsHarvesting Machine
of pure fat tissuesHarvesting Machine
of pure fat tissues
• Trial of massive culturing of stem cells through 3 dimensional cell culturing technology. • Introduction of stem cell culturing technology to stem cell extractor is under process. • With fully automated system of stem cell isolation and culture, our advanced technology can be applied for stem cell production for cell therapy.
Fat Transfer Device integrating liposuction and centrifugation in one system Fast & Massive Fat Transfer can be performed.Can harvest pure, concentrated fat tissues using
closed-circuit AFT syringes equipped with weight-mesh piston to remove free oils and prevent contamination.
KFDA Approved (2004) w/ proprietary patentSales Contract with Biomaster in JapanDomestic Sale to Plastic & Cosmetic SurgeonsDomestic(15) and overseas(11) patent protected
• In 2006, development of automated stem cell extractor.• Can automatically isolate stem cells from fat tissues at fast speed. •Can separate stem cell with high yield rate at 37℃ .•All processes are automatically made.• 1st Quality test was finished w/ enhancing its performance. •Pending Patent (#20-2004-0010685) –Centrifuging unit integrating liposuction and fat injection. And other patent (Domestic 9, Overseas 9)
Automated Stem Cell Extractor (YC-100) Cell Therapy System
2001 2004 2007 2010
01. 01. Bio MasterBio Master의의 개요개요Patents for Automated Stem Cell Extractor(YC-100)
Centrifuging Device
Centrifuging Device & MethodCentrifuging Device & Method
Application No. : 10-2006-0111264, Applied Date : Nov. 10, 2006Summary: Device & Method featured by angle control part to control a receptor angle so that collected top layer fluids during centrifugation can flow down at over 90 angle.
Application No. : 10-2006-0060513, Applied Date: June 30, 2006Summary : Centrifugal device having a conical shape to move up materials on the receptor to the top layerby centrifugal force and also having a filter on the top layer of the receptor to pass only some parts of thematerials through.
Centrifuging Device & MethodCentrifuging Device & Method
Centrifuging Device & MethodCentrifuging Device & Method
Application No. : 10-2006-0111266, Applied Date : Nov. 10, 2006Summary: Centrifugal device and method to distinguish and separate fluids by differences of large or small 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.
Application No. : 10-2006-0111265, Applied Date : Nov. 10, 2006Summary: Centrifugal device & Method to exactly and easily distinguish and recover fluid layers by difference of specific gravities and fluid resistance between the separated fluid materialswhen opening part of the container is opened after revolution body is revolved by centrifugation.
01. 01. Bio MasterBio Master의의 개요개요Technological Value of Stem cell Extractor, YC-100(International Search Report)
CYTORI THERAPEUTICS TISSUE GENESIS
MEDIKAN YC-100
01. 01. Bio MasterBio Master의의 개요개요Automated Stem Cell Extraction & Culture Device (YC-100)
Basic SpecificationBasic SpecificationBasic Specification External Design of YC-100 External Design of YCExternal Design of YC--100 100
구 분
Items Specifications
Max RPM 3,000 rpm
Max Capacity 2,000 ml
Max centrifugal force >200g
Rotor radius of rotation range 0.1 rpm
Drive system Ultra-precision Servo Drive control
Speed preset and display 1 rpm
Temperature Range -20 ℃ ~ +50 ℃
Timer 99 hrs 59 min 59 secAnd “Hold” for free run
Acceleration Time 3 stage
Deceleration Time 10 setting
Programmability 100 memory
Digital Display RPM, RCF, Time, Temperature, Temp, Limit, Program, Acel/Decel
Power Requirements 220 V, 60 Hz, single phase, 3.0 kw
Dimension 1310 W×620 D×1230 H / mm
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.
01. 01. Bio MasterBio Master의의 개요개요
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
Automated Stem Cell Extraction & Culture Device (YC-100)
01. 01. Bio MasterBio Master의의 개요개요Automated Stem Cell Extraction & Culture Device (YC-100)
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.
01. 01. Bio MasterBio Master의의 개요개요
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.
Automated Stem Cell Extraction & Culture Device (YC-100)
01. 01. Bio MasterBio Master의의 개요개요Automated Stem Cell Extraction & Culture Device (YC-100)
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.
01. 01. Bio MasterBio Master의의 개요개요
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
Automated Stem Cell Extraction & Culture System, (YC-100)
Stroboscope Light
Moving Image Camera
01. 01. Bio MasterBio Master의의 개요개요Automated Stem Cell Extraction & Culture System, (YC-100)
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
01. 01. Bio MasterBio Master의의 개요개요Efficiency Test for Stem Cell extraction & Culture System, YC-100
Harvesting fat tissues by Lipokit (made by Medikhan Corp.)
To minimize errors, efficiency test with only highly concentrated fat tissues having over 90% purity to be harvested by Lipokit
vs.
Manual Method: manual isolation of stem cells from fat tissues directly by skilled expert
Automated Method: isolation of stem cell by YC-100
Collagenase treatment
Separation of fat tissues by centrifuge
Washing by PBS buffer
Cell harvest
Cell counting
Comparison & analysis of efficiency rates of stem cells isolated by experts and by YC-100
Analysis of over 3 times per session using same fat tissues
Testing totally over 20 sessions of efficiency rates
Counting the number of total cells (ADSCs & fibrobast etc.) isolated from fat tissues by automatic cell counter after staining with PI (Propidium Iodide)
01. 01. Bio MasterBio Master의의 개요개요
Manual Automated
Cell number(0.1x million cells/fat1cc)
9.65 10.9
Yield (%) 100 112.84
0
20
40
60
80
100
120
140
수동 자동
Isolation method
Isol
atio
n yi
eld
(%)
Efficiency Test of Stem Cell Isolation by YC-100
manual Automated
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.
01. 01. Bio MasterBio Master의의 개요개요Processing for Isolation of Adipose-derived stem cell (a few processes are intentionally omitted. Confidential.)
01. 01. Bio MasterBio Master의의 개요개요Development of Culture Technology
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.
x200
0
5
10
15
20
25
30
35
40
45
50
initial 6h 3d 6d 10d
Culture time
Cell
num
ber
(1X
105
cells
/bea
ds 1
cc) PLLA beads
Fat scaffold
Dextran beads
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
10-2005-0078382 : As an injection material, solid chitosan powder with sponge structure to be injected for human body and its production method
10-2006-0079725 : Micro Scaffolds with non-spherical 3 dimensional stereo structure to be produced by rapid prototyper and used for cell culture and transfer.
10-2007-0041058 : Cell culturing scaffold with increased specific gravity and its production method.
< In vitro proliferation test using ADSC P0 >
Development of 3 Dimensional scaffolds for cell proliferation
01. 01. Bio MasterBio Master의의 개요개요Stem Cell Culturing System
<Simulation for 3 Dimensional Stem Cell Culture >
01. 01. Bio MasterBio Master의의 개요개요
Dreaming of Future High-tech Bio Business
MedikhanDevelopment of automated Stem Cell Extraction & Culture Device
Exploitation of Cell TherapyCell Banking Business
Biomaster (Japan)
Clinical Research using Stem Cells
(Extraction of Stem Cell + Bioreactor + Cell therapy)
Potential Prospect of YC-100 Stem Cell Extraction and Culture Device