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Novel Platform Technology

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Novel Platform Technology for Scale Novel Platform Technol Platform Technology Novel Novel Platform Technology for Scale Novel Platform Technology for Scale for Scale Down and Optimisation for Optimisation for Process Optimization Down and Process Down and Process Down and Process Opt Down and Process Optimisation for for Regenerative Medicine Regenerative Medicine Regenerative Medicine Regenerative Medicine Regenerative Medicine is now part of the Sartorius Stedim Biotech Group is now part of the Sartorius Stedim Biotech Group JoAnne Swihart, Kim Bure, Rosemary Drake and Barney Zoro JoAnne Swihart, Kim Bure, Rosemary Drake and Barney ZoroSwihart, Kim Bure, Rosemary Drake and Barney Zoro Swihart, Kim Bure, Rosemary Drake and Barney Zoro JoAnne JoAnne JoAnne Swihart, Kim Bure, Rosemary Drake and Barney Zoro Summary Summary Summary ambr15 15 Summary ambr15TM ambr® ambr15 Parallel automated • A significant challenge for the Regenerative Medicine (RM) industry is t Parallel automated • A significant challenge for the Regenerative Medicine (RM) industry is to develop cell • A significant challenge for the Regenerative Medicine (RM) industry is to develop cell - A significant challenge for the Regenerative Medicine (RM) industry is to develop cell culture Parallelautomated automated Parallel • A significant challenge for the Regenerative Medicine (RM) industry is to develop cell are affordable, well characterised and can be processes for are affordable, well characterised and can be s culture processes affordable,affordable, well characterised and culture processesensure culture scaled up that processes that are that are well characterised and can be scaled upcan production to that for be scaled up for micro-scale micro-scale culture processes that clinical and commercial success. and can be scaledensure clinical micro-scale bioreactor are affordable, well characterised up for micro-scale production to ensure success. production to and commercial success. production to ensure clinical and commercial success. clinical and commercial production to ensure clinical and commercial success. bioreactor system bioreactor system system bioreactor system • ambr15 is an automated micro-scale bioreactor system that mimics th • ambr15 is an automated micro-scale bioreactor system that mimics the features and • ambr15 is an automated micro-scale bioreactor system that mimics the features and - ambr® 15 is an automated micro-scale bioreactor system that mimics the features and process • ambr15 is an automated micro-scale bioreactor system that mimics the features and Summary process control (pH, DO, temperature, stirring rate) provided by much larger scale a process control (pH, DO, temperature, stirring rate) provided by much larger scale 3 process control (pH, DO, temperature, stirring rate) provided by much control (pH, DO, temperature, stirring rate) provided by much larger scale bioreactors, but in 7 3 4 4 process control (pH, DO, temperature, stirring rate) provided by much larger scale high 3 4 bioreactors, but in a volume of 10-15ml. Parallel processing capability and excellent bioreactors, but in a volume of 10-15ml. Parallel processing capability and excellent bioreactors, but in a volume of 10-15ml. Parallel processing capability a volume of 10 - 15 mL. Parallel processing capability and excellent consistency enable rapid, bioreactors, but in a volume of 10-15ml. Parallel processing capability and excellent consistency process improvement and optimisation, including DoE studies.and optimisation, consistency enable rapid, high throughput process improvement and optimisation, consistency enable rapid, high throughput process improvement and o throughput enable rapid, high throughput process improvement 7 consistency enable rapid, high throughput process improvement and optimisation, 7 including is widely used including DoE studies. • 24 – 48 parallel stirred, sparged miniature including DoE studies. • 24 – 48 parallel stirred, sparged miniature - 24 – 48 parallel stirred, sparged miniature - ambr® 15 DoE studies. including DoE studies. in the bio-process industry for process development, cell line selection and • 24 – 48 parallel stirred, sparged miniature bioreactors bioreactors bioreactors. • ambr15 is widely used in the bio-process industry for process development, cell line • ambr15 is widely used in the bio-process industry for process developm effective media optimisation in less time with reduced reagent use• ambr15 is widely used in the bio-process industry for process development, cell line and labour saving (1, 2). It can bioreactors are disposable for ease of use and • Bioreactors are disposable for ease of use - Bioreactors • ambr15 is widely used in the bio-process industry for process development, cell line media• Bioreactors are disposable for ease of use selection and effective media and rapid throughputless time with reduced re • Bioreactors are disposable for ease of use and rapid throughput rapid throughput. selection and effectivesmall scale continuous culture perfusion mimic. selection and effective optimisation in less time with reduced reagent use and optimisation in also be used as a novel media optimisation in less time with reduced reagent use and and rapid throughput selection and effective media optimisation in less time with reduced reagent use and ambr® 15 workstation ambr15 workstation amb • DO and pH sensing allows control of • DO and pH sensing allows control of - DO and pH sensing allows control of individual 6 8 labour saving (1, 2). It can also be used as a novel small scale continuous culture cell labour saving (1, 2). It can also be used as a novel small scale continuous culture labour saving (1, 2). It can also be used as a novel small scale continuo ambr15 workstation • DO and pH sensing allows control of - ambr® 15 has been successfully used in RM applications; including processing microcarriers, individual bioreactors individual bioreactors bioreactors. labour saving (1, 2). It can also be used as a novel small scale continuous culture perfusion mimic. perfusion mimic. perfusion mimic. individual bioreactors enables automated liquid • Liquid handling robot enables automated • Liquid handling robot enables automated - Liquid handling robot ambr vessel aggregates e.g. embryoid bodies and HSCs (3, 4). 6 9 perfusion mimic. • Liquid handling robot enables automated liquid additions and sampling, reducing full liquid additions and sampling, reducing full 10 - 8 mL fill 15 6 additions and sampling, reducing full time ambr15 vessel 1 Automated Liquid Handler • ambr15 has been successfully used in RM applications; including processing • ambr15 has been successfully used in RM applications; including processing • ambr15 has been successfully used in RM applications; including proc 8 liquid additions and sampling, reducing full volume time employee requirement time employee requirement employee requirement. - High throughput tools with parallel processing, such as ambr® 15, help address a major ambr15 vessel 1 Automated Liquid Handler 10-15 mL fill • ambr15 has been successfully used in RM applications; including processing 2 Culture Stations time employee requirement software. microcarriers, cell aggregates e.g. embryoid bodies and HSCs (3, 4).resources and microcarriers, cell aggregates e.g. embryoid bodies and HSCs (3, 4). microcarriers, cell aggregates e.g. embryoid bodies and HSCs (3, 4). • System controlled by central software • System controlled by central software 10-15 mL fill - System controlled by central volume 2 Culture Stations (12 vessels each) biomanufacturing bottleneck and improve economic models by reducing the 9 microcarriers, cell aggregates e.g. embryoid bodies and HSCs (3, 4). • System controlled by central software • Fully automated, parallel bioreactors • Fully automated, parallel bioreactors volume 3 (12 vessels each) Sample plates - Fully automated, parallel bioreactors enable 1 Automated Liquid Handler 6 Sparge tube 9 • High throughput tools with parallel processing, such as ambr15, help address a major • Fully automated, parallel bioreactors for • High throughput tools with parallel processing, such as ambr15, help a • High throughput tools with parallel processing, such as ambr15, help address a major timescale needed to develop robust and cost-effective manufacturing processes, so bringing novel enable simple implementation of DoE rapid enable simple implementation of DoE 3 Sample plates 4 Liquid additions simple implementation of DoE studies 2 Culture Stations 7 Sample port • High throughput tools with parallel processing, such as ambr15, help address a major enable simple implementation of DoE 4 Tips bottleneck and improve economic models by reduci 5 resources studies for rapid process development and reducing theLiquid additions studies for rapid process development and (12 vessels each) 8 Impeller biomanufacturing bottleneck and improve economic models by reducing the resources and improve economic models by biomanufacturing bottleneck studies for rapid process development and biomanufacturing therapies to patients sooner. process development and characterisation. 5 Tips 6 Sparge tube biomanufacturing bottleneck and improve economic models by reducing the resources characterisation characterisation 3 Sample plates 9 pH and DO sensing - Fits into a biosafety cabinet to maintain sterility. and timescale needed to develop robust and cost-effective manufacturing processes, and timescale needed to develop robust and cost-effective manufacturing processes, and timescale needed to develop robust and cost-effective manufactu 6 additions Sample port 7 Sparge tube characterisation 4 Liquid • Fits into a biosafety cabinet to maintain • Fits into a biosafety cabinet to maintain and timescale needed to develop robust and cost-effective manufacturing processes, 8 Impeller 5 Tips 7 Sample port • Fits into a biosafety cabinet to maintain so bringing novel therapies to patients sooner. so bringing novel therapies to patients sooner. so bringing novel therapies tosterility patients sooner. sterility 8 Impeller 9 pH and DO sensing so bringing novel therapies to patients sooner. sterility 9 pH and DO sensing Inoculum Density (x105 cells | m L) Inoculum Density (x10 cells | m L) 5 Static Control EB size (72h) (µm) 194 ± 192 256 ± 57 232 ± 57 Figure 2. P17-7 cells were cultured in hES media with 20ng/ml of FGF-2 and 10µM Figure 1. NK92 cells were adapted to grow in microscale bioreactors from static cultures Figure 2. P17-7 cells were cultured in hES media with 20ng/ml of FGF-2 and 10µM ROCKi. EBs were formed by 24 hours on an orbital shaker at 70rpm and then by testing different agitation rates and inoculum densities. The microscale bioreactors ROCKi. EBs were formed by 24 hours on an orbital shaker at 70rpm and then ROCKi. Figure 2. P17-7 cells were cultured in hES media with 20 ng | mL of FGF-2 and 10μM transferred to ambr15 vessels at 5 x 105 cells/ml, and cultured on the ambr15 platform were able to maintain a uniform cell suspension throughout the experiment. (A) Fold transferred to ambr15 vessels at 5 x 105 cells/ml, and cultured on the ambr15 platform EBs were formed by 24 hours on an orbital shaker at 70rpm and for up to 96 hours at 300rpm (A and C) or 600rpm (B and D). then transferred to ambr® 15 expansion and (B) cell viability of NK92 cells on day 4 across seeding cell densities for up to 96 hours at 300rpm (A and C) or 600rpm (B and D). vessels at 5 x 105 cells | mL, and cultured on the ambr® 15 platform for up to 96 hours at (2-7x105 cells/mL) and 4 different agitation rates (450 and 1100 rpm). (C) Representative 300 rpm (A and C) or 600 rpm (B and D). micrographs of NK92 cells in static culture (left) compared to microscale bioreactors (right) (scale bar 100mm). Figure 3. NKx2-5 and hES2 cells were seeded and formed aggregates in an orbital shaker Figure 2. P17-7 cells were cultured in hES media with 20ng/ml of FGF-2 and 10µM Figure 1. NK92 cells were adapted to grow in microscale bioreactors from static cultures Figure 3. NKx2-5 and hES2 cells were seeded and formed aggregates in an orbital shaker for 24 hrs, before being transferred in duplicate to the ambr15 platform or remaining in ROCKi. EBs were formed by 24 hours on an orbital shaker at 70rpm and then by testing different agitation rates and inoculum densities. The microscale bioreactors 24 Figure 3. NKx2-5 and hES2 cells were seeded and formed aggregates in an orbital shaker for for 24 hrs, before being transferred in duplicate to the ambr15 platform or remaining in the orbital shaker. On D14 (A) viable cell counts were used to calculate the total cell fold transferred to ambr15 vessels at 5 x 105 cells/ml, and cultured on the ambr15 platform were able to maintain a uniform cell suspension throughout the experiment. (A) Fold hrs, before being transferred in duplicate to the ambr® 15 platform or remaining in the orbital the orbital shaker. On D14 (A) viable cell counts were used to calculate the total cell fold expansion and (B) cTnT positive cells were quantified to calculate cardiomyocyte output. for up to 96 hours at 300rpm (A and C) or 600rpm (B and D). cell fold expansion and expansion and (B) cell viability of NK92 cells on day 4 across seeding cell densities shaker. On D14 (A) viable cell counts were used to calculate the total expansion and (B) cTnT positive cells were quantified to calculate cardiomyocyte output. (C) Representative micrographs of NKx2-5 and hES2 aggregates in orbital shaker culture (2-7x105 cells/mL) and 4 different agitation rates (450 and 1100 rpm). (C) Representative (B) cTnT positive cells were quantified to calculate cardiomyocyte output. (C) Representative (C) Representative micrographs of NKx2-5 and hES2 aggregates in orbital shaker culture (left) compared to microscale bioreactors (right). shaker culture (left) compared to micrographs of NK92 cells in static culture (left) compared to microscale bioreactors micrographs of NKx2-5 and hES2 aggregates in orbital (left) compared to microscale bioreactors (right). (right) (scale bar 100mm). microscale bioreactors (right). Data courtesy of: Data courtesy of: Data courtesy of: Data courtesy of: Reliable automation of Proven high throughput Proven high throughput Proven high throughput tool for microcarrier studies tool for scale up scale scale up tool forUp ambr15 can be effectively used as a high throughput scale-down model for Industrial users have demonstrated that the ambr15 system matches the Industrial users Industrial users have demonstrated that the ambr15 system matches the microcarrier work. have demonstrated that the ambr® 15 system matches the data produced in much larger, for example 5 L, bench-scale systems data produced in much larger, for example 5 L, bench-scale systems. data produced in much larger, for example 5 L, bench-scale systems • Tunable liquid handling offers consistent microcarrier seeding (A) Cell Growth • Settling method (B) enables challenging media exchange operations • Cell growth on microcarriers is comparable to standard culture a) Reliable automation of optimization Proven high and media optimisation Prove DoE process and media optimisation DoE pr DoEprocess and media process throughput DoE for scale up microcarrier has been successfully implemented by a number of industrial The ambr15 p tool 15 platform studies tool fo The ambr15 platform has been successfully implemented by a number of The ambr® The ambr15 platform has been successfully implemented by a number of industrial companies for media development and feed strategy optimisation. companies for media development and feed strategy optimization. ambr15 can be effectively used as a high throughput scale-down model for Industrial users have demonstrated that the ambr15 system matches the industrial companies for media development and feed strategy optimisation. microcarrier work. data produced in much larger, for example 5 L, bench-scale systems The ability to compare multiple variables simultaneously in one run provides: The ability to compare multiple variables simultaneously in one run provides: The ability to compare multiple variables simultaneously in one run provides: • - A many-fold data production increase in less time and a smaller footprint A many-fold data production increase in less time and a smaller footprint • Tunable liquid handling offers consistent microcarrier seeding (A) • A many-fold data production increase in less time and a smaller footprint • - Higher comparability for DOE run in one experiment Higher comparability for DOE run in one experiment • Settling method (B) enables challenging media exchange operations • Higher comparability for DOE run in one experiment • - Greater process understanding for manufacturing and FDA filing Greater process understanding for manufacturing and FDA filing • Cell growth on microcarriers is comparable to standard culture • Greater process understanding for manufacturing and FDA filing a) 3 Rounds of Feed Optimization Product Concentration Platform industrial com Industrial u data produ The ability to • A many-fol • Higher com • Greater pro Figure 3. NKx Figure 2. P17for 24 hrs, bef ROCKi. EBs w the orbital sha transferred to expansion and for up to 96 h (C) Represen (left) compare Data courtesy of: Data courtesy of: Data courtesy of: Data courtesy of: Data courtesy of: ambr® 15 can be ambr15 can be effectively used as a high throughput scale-down model for microcarrier work. effectively used as a high throughput scale-down model for microcarrier work. microcarrier work. • Tunable liquid handling offers consistent microcarrier seeding (A) - Tunable liquid handling offers consistent microcarrier seeding (A). • Tunable liquid handling offers consistent microcarrier seeding (A) • Settling method (B) enables challenging media exchange operations - Settling method (B) enables challenging media exchange operations. • Settling method (B) enables challenging media exchange operations • Cell growth on microcarriers is comparable to standard culture - Cell growth on microcarriers is comparable to standard culture. • Cell growth on microcarriers is comparable to standard culture 100 a) 90 a) a) Data courtesy of: Data courtesy of: Data courtesy of: Reliable automation of Reliable automation Reliable automation ofof microcarrier microcarrier studies studies microcarrier studies ambr15 can be effectively used as a high throughput scale-down model for Figure 1. NK92 cells were adapted to grow in microscale bioreactors from static cultures Figure 1. NK92 cells were adapted to grow in microscale bioreactors from static cultures Figure 1. NK92 cells were adapted to grow in microscale bioreactors from static cultures by testing by testing different agitation rates and inoculum densities. The microscale bioreactors by testing different agitation rates and inoculum densities. The microscale bioreactors different agitation rates and inoculum densities. The microscale bioreactors were able to maintain a were able to maintain a uniform cell suspension throughout the experiment. (A) Fold were able to maintain a uniform cell suspension throughout the experiment. (A) Fold of NK92 uniform cell suspension throughout the experiment. (A) Fold expansion and (B) cell viability expansion and (B) cell viability of NK92 cells on day 4 across seeding cell densities expansion and (B) cell viability of NK92 cells on day 4 across seeding cell densities (450 cells on day 4 across seeding cell densities (2-7x105 cells | mL) and 4 different agitation rates (2-7x105 cells/mL) and 4 different agitation rates (450 and 1100 rpm). (C) Representative (2-7x105 cells/mL) and 4 different agitation rates (450 and 1100 rpm). (C) Representative and 1100 rpm). (C) Representative micrographs of NK92 cells in static culture (left) compared micrographs of NK92 cells in static culture (left) compared to microscale bioreactors to micrographs of NK92 cells in static culture (left) compared to microscale bioreactors microscale bioreactors (right) (right) (scale bar 100mm). (scale bar 100 mm). (right) (scale bar 100mm). Cardiomyocytes per hESC input Figure 5. A CHO cell line producing a large therapeutic protein was grown under proprietary conditions. The error bars represent the acceptance criteria for matching different scale bioreactors (±20%). Data provided courtesy of Stanislas Augusto, Sanofi Aventis. Figure 6. a) Data courtesy of CobraBio, Feed and media screening including DoE resulted in a 7-fold increase in titer over 2 months. b) A DoE mixture design identifies of optimal growth media, operating in a perfusion mimic mode. Figure 4. a) Cytodex1 microcarriers were c) c) automatically added to 6 ambr® 15 vessels, achieving CV < 1% across the 6 vessels. b) Culture is automatically sampled and settled in the pipette tip. 0 2 4 6 8 10 12 14 16 18 20 Microcarrier are dispensed back to the Time (days) culture and the media is discarded. c) Vero cells (ATCC) achieved confluence ambr 15 ambr 15 5 L BIOSTAT® B on the microcarriers. Figure 6. a) Data courtesy of CobraBio, Feed and media screening including DoE Figure 5. A CHO cell line producing a large therapeutic protein was grown under Figure 5. A CHO cell line producing a large therapeutic protein was grown under Figure 4. a) Cytodex1 microcarriers were automatically added to 6 ambr15 vessels, achieving Figure 4. a) Cytodex1 microcarriers were automatically added to 6 ambr15 vessels, achieving Figure 4. a) Cytodex1 microcarriers were automatically added to 6 ambr15 vessels, achieving Figure 6. a) Data courtesy of CobraBio, Feed and media screening including DoE Figure 5. A CHO cell line producing a large therapeutic protein was grown under Figure 4. a) Cytodex1 microcarriers were automatically added to 6 ambr15 vessels, achieving resulted in a 7-fold increase in titer over 2 months. b) A DoE mixture design identifies proprietary conditions. The error bars represent the acceptance criteria for matching proprietary conditions. The error bars represent the acceptance criteria for matching CV < 1% across the 6 vessels. b) Culture is automatically sampled and settled in the pipette CV < 1% across the 6 vessels. b) Culture is automatically sampled and settled in the pipette CV < 1% across the 6 vessels. b) Culture is automatically sampled and settled in the pipette resulted in a 7-fold increase in titer over 2 months. b) A DoE mixture design identifies proprietary conditions. The error bars represent the acceptance criteria for matching CV < 1% across the 6 vessels. b) Culture is automatically sampled and settled in the pipette Bibliography of optimal growth media, operating in a perfusion mimic mode. different scale bioreactors (±20%). Data provided courtesy of Stanislas Augusto, Sanofi different scale bioreactors (±20%). Data provided courtesy of Stanislas Augusto, Sanofi tip. Microcarrier are dispensed back to the culture and the media is discarded. c) Vero cells tip. Microcarrier are dispensed back to the culture and the media is discarded. c) Vero cells tip. Microcarrier are dispensed back to the culture and the media is discarded. c) Vero cells Phone +44.1763.227200 1. Hsu WT., et. al. Advanced microscale bioreactor system: a representative scale-down model for of optimal growth media, operating in a perfusion mimic mode. different scale bioreactors (±20%). Data provided courtesy of Stanislas Augusto, Sanofi tip. Microcarrier are dispensed back to the culture and the media is discarded. c) Vero cells bench-top bioreactors. Cytotechnology (2012). Aventis Aventis (ATCC) achieved confluence on the microcarriers. (ATCC) achieved confluence on the microcarriers. Sartorius Stedim Biotech UK (ATCC) achieved confluence on the microcarriers. 2. oses S. et al. Assessment of AMBRTM as a model M York Way, Royston, Fax +44.1763.227201 Aventis (ATCC) achieved confluence on the microcarriers. for high-throughput cell culture process development strategy. Advances in Biosceince and hES cells s • ambr15 sig • cardiomyoc The agitatio C) giving la control at 7 Fold Expansion Cellular aggre cultured in am successfully e hESC e Embryo in amb culture Cellular aggregates derived from NKx2-5 and hES2 cells can be successfully Cellular aggregates such as embryoid bodies (EB) can be successfully The ambr15 system can be used for systematic investigation of critical Cellular aggregates derived from NKx2-5 and hES2 cells can be Cellular aggregates derived from NKx2-5 and hES2 cells can be expanded and differentiated in ambr® 15. cultured in ambr15. process parameters, with automatic feeding, control and sampling. successfully expanded and differentiated in ambr15. successfully expanded and differentiated in ambr15. - ambr® 15 significantly improved aggregate expansion (A) and cardiomyocyte • • ambr15 significantly improved aggregate expansion (A) and Optimum stirrer speed for NK92 culture found to be 450 RPM • hES cells show good survival, and proliferate • • cardiomyocyte differentiation (B) compared to orbital shaker control. differentiation (B) compared to orbital shaker control. • ambr15 significantly improved aggregate expansion (A) and The agitation speed influences cell yield, with a speed of 300rpm (A and No strong correlation between inoculum density and D4 viability cardiomyocyte differentiation (B) compared to orbital shaker control. • NK92 cells in ambr15 are a uniform, well mixed cell suspension C) giving larger EB size than 600rpm (B and D) and the orbital shaker control at 70rpm Expansion in Orbital Shaker vs. ambr 15 hESC Cardiomyctes per hESC input Cellular aggregates such as embryoid bodies (EB) can be successfully cultured Cellular aggregates such as embryoid bodies (EB) can be successfully in The ambr15 system can be used for systematic investigation of critical Cellular aggregates such as embryoid bodies (EB) can be successfully ambr® 15. cultured in ambr15. process parameters, with automatic feeding, control and sampling. cultured in ambr15. hES cells show good survival, and proliferate • - Optimum stirrer speed for NK92 culture found to be 450 RPM • hES cells show good survival, and proliferate • • The agitation speed influences cell yield, with a speed of 300rpm (A and hES cells show good survival, and proliferate speed of 300 rpm (A and • - No strong correlation between inoculum density and D4 viability C) The agitation speed influences cell yield, with a • • C) giving larger EB size than 600rpm (B and D) and the orbital shaker The agitation speed influences cell yield, with a speed of 300rpm (A and NK92 cells in ambr15 are a uniform, well mixed cell suspension giving larger EB size than 600 rpm (B and D) and the orbital shaker control at C) giving larger EB size than 600rpm (B and D) and the orbital shaker control at 70rpm 70 rpm control at 70rpm a) Embryoid bodies and differentiation hESC expansion and differentiation Optimizing culture conditions hESC expansion successfully hESC expansion and differentiation cultured in ambr15 in ambr15 for NK92 cells in ambr® in ambr15 15 Fold Expansion The ambr® 15 system can be used for systematic investigation of critical process The ambr15 system can be used for systematic investigation of critical The ambr15 system can be used for systematic investigation of critical parameters, with automatic feeding, control and sampling. process parameters, with automatic feeding, control and sampling. process parameters, with automatic feeding, control and sampling. - Optimum stirrer speed for NK92 culture found to be 450 rpm. • Optimum stirrer speed for NK92 culture found to be 450 RPM • Optimum stirrer speed for NK92 culture found to be 450 RPM • No strong correlation between inoculum density and D4 viability - No strong correlation between inoculum density and D4 viability. • No strong correlation between inoculum density and D4 viability • NK92 cells in ambr15 are a uniform, well mixed cell suspension - NK92 cells in ambr® 15 are a uniform, well mixed cell suspension. • NK92 cells in ambr15 are a uniform, well mixed cell suspension a) a) a) Embryoid bodies successfully Optimizingbodies successfully culture conditions Embryoid bodies successfully Embryoid cultured in ambr15 for NK92 cells cultured in ambr® cultured in ambr15 15 Fold Expansion Optimizing culture conditions Optimizing culture conditions culture conditions Optimizing for NK92 cells for NK92 cells for NK92 cells Figure 6. Figure 5. A CH resulted i proprietary co of optima different scale Aventis Biotechnology (2012). Herts, SG8 5WY www.tapbiosystems.com TAP Biosystems BibliographyE. et. al., A novel automated bioreactor for scalable process optimisation of haematopoietic stem cell culture. J. Biotechnology (2012). Bibliography Bibliography 3. Ratcliff TAP Biosystems - a Sartorius Group Company TAP Biosystems - a Sartorius Group Compa www.sartorius.com 1. su WT., et. al. Advanced microscale bioreactor system: a representative scale-down model for bench-top bioreactors. Cytotechnology (2012). H len KE. 1. su WT., et. al. Advanced microscale bioreactor system: a representative scale-down model for bench-top bioreactors. Cytotechnology (2012). H 1. su WT., et. al. Advanced microscale bioreactor system: a representative scale-down model for bench-top bioreact H is now part of the Sartorius Stedim Biotech Group Bibliography et al. Production of erythorocytes from directly isolated or Delta1 Notch ligand expanded CD34+ hematopoietic progenitor cells: process 4. G TAP Biosystems - a Sartorius Group Company M M 2. su WT., et. al. Advanced microscale bioreactor system: a representative scale-down model for bench-top bioreactors. Cytotechnology (2012). 2. oses S. et al. Assessment of AMBRTM as a model for high-throughput cell culture process development strategy. Advances in Biosceince and 1. M oses S. et al. Assessment of AMBRTM as a model for high-throughput cell culture process development strategy. Advances in Biosceince and H characterization, monitoring and implications for manufacture. Cytotherapy (2013). Europe & Rest of World 2. oses S. et al. Assessment of AMBRTM as a model for high-throughput cell culture process development strategy. Europe & Rest of World Biotechnology (2012). Biotechnology (2012). M oses S. et al. Assessment of AMBRTM as a model for high-throughput cell culture process development strategy. Advances in Biosceince and 2. Biotechnology (2012). Europe & Rest of World TAP Biosystems Telephone: TAP Biosystems Telepho 3. atcliff E. et. al., A novel automated bioreactor for scalable process optimisation of haematopoietic stem cell culture. J. Biotechnology (2012). R 3. atcliff E. et. al., A novel automated bioreactor for scalable process optimisation of haematopoietic stem cell culture. J. Biotechnology (2012). R 3. atcliff E. et. al., A novel automated bioreactor for scalable process optimisation of haematopoietic stem cell cultur R Biotechnology (2012). TAP Biosystems Telephone: Publication No. : SBI1550-e151001 Order No. : 85037-552-63 York Way, Royston +44 (0)1763 227200 York Way, Royston +44 (0)17 4. 4. len KE. et al. Production of erythorocytes from directly isolated or Delta1 Notch ligand expanded CD34+ hematopoietic progenitor cells: process G 4. len KE. et al. Production of erythorocytes from directly isolated or Delta1 Notch ligand expanded CD34+ hematopo G 3. G len KE. et al. Production of erythorocytes from directly isolated or Delta1 Notch ligand expanded CD34+ hematopoietic progenitor cells: process R atcliff E. et. al., A novel automated bioreactor for scalable process optimisation of haematopoietic stem cell culture. J. Biotechnology (2012). York Way, Royston +44 (0)1763 227200 Herts SG8 5WY Fax: Herts SG8 5WY Fax: characterization, monitoring and implications for manufacture. Cytotherapy (2013). characterization, monitoring and implications for manufacture. Cytotherapy (2013). 4. characterization, monitoring and implications for manufacture. Cytotherapy (2013). G len KE. et al. Production of erythorocytes from directly isolated or Delta1 Notch ligand

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