PL2488635T3 - Proces do oczyszczania adenowirusa z hodowli o dużej gęstości komórek - Google Patents
Proces do oczyszczania adenowirusa z hodowli o dużej gęstości komórekInfo
- Publication number
- PL2488635T3 PL2488635T3 PL10771402T PL10771402T PL2488635T3 PL 2488635 T3 PL2488635 T3 PL 2488635T3 PL 10771402 T PL10771402 T PL 10771402T PL 10771402 T PL10771402 T PL 10771402T PL 2488635 T3 PL2488635 T3 PL 2488635T3
- Authority
- PL
- Poland
- Prior art keywords
- cell density
- high cell
- density cultures
- adenovirus purification
- adenovirus
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
- C12N15/86—Viral vectors
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/10011—Adenoviridae
- C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
- C12N2710/10341—Use of virus, viral particle or viral elements as a vector
- C12N2710/10343—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/10011—Adenoviridae
- C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
- C12N2710/10351—Methods of production or purification of viral material
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Plant Pathology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US27906009P | 2009-10-15 | 2009-10-15 | |
| EP09173119 | 2009-10-15 | ||
| PCT/EP2010/065436 WO2011045381A1 (en) | 2009-10-15 | 2010-10-14 | Process for adenovirus purification from high cell density cultures |
| EP10771402.4A EP2488635B1 (en) | 2009-10-15 | 2010-10-14 | Process for adenovirus purification from high cell density cultures |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| PL2488635T3 true PL2488635T3 (pl) | 2014-04-30 |
Family
ID=41563777
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PL10771402T PL2488635T3 (pl) | 2009-10-15 | 2010-10-14 | Proces do oczyszczania adenowirusa z hodowli o dużej gęstości komórek |
Country Status (16)
| Country | Link |
|---|---|
| US (1) | US8470585B2 (pl) |
| EP (1) | EP2488635B1 (pl) |
| JP (1) | JP5465331B2 (pl) |
| KR (1) | KR101805938B1 (pl) |
| CN (1) | CN102575233B (pl) |
| AU (1) | AU2010305768B2 (pl) |
| BR (1) | BR112012008507B8 (pl) |
| CA (1) | CA2777116C (pl) |
| DK (1) | DK2488635T3 (pl) |
| EA (1) | EA028875B1 (pl) |
| ES (1) | ES2445713T3 (pl) |
| HK (1) | HK1174949A1 (pl) |
| MX (1) | MX2012004221A (pl) |
| PL (1) | PL2488635T3 (pl) |
| WO (1) | WO2011045381A1 (pl) |
| ZA (1) | ZA201202524B (pl) |
Families Citing this family (39)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| PL2350268T3 (pl) | 2008-11-03 | 2015-05-29 | Crucell Holland Bv | Sposób wytwarzania wektorów adenowirusowych |
| SG11201405228VA (en) | 2012-03-12 | 2014-11-27 | Crucell Holland Bv | Batches of recombinant adenovirus with altered terminal ends |
| US8932607B2 (en) | 2012-03-12 | 2015-01-13 | Crucell Holland B.V. | Batches of recombinant adenovirus with altered terminal ends |
| US9125870B2 (en) | 2012-03-22 | 2015-09-08 | Crucell Holland B.V. | Vaccine against RSV |
| EP2827895B1 (en) | 2012-03-22 | 2017-08-09 | Janssen Vaccines & Prevention B.V. | Vaccine against rsv |
| WO2013154928A1 (en) * | 2012-04-08 | 2013-10-17 | Kapre Subhash V | Systems and methods of virus propagation in cell culture for vaccine manufacture |
| CA2902650A1 (en) | 2013-02-28 | 2014-09-04 | Psioxus Therapeutics Limited | A process for the production of adenovirus |
| DK3467108T3 (da) | 2013-03-14 | 2024-06-10 | Translate Bio Inc | Fremgangsmåder til oprensning af messenger-RNA |
| AP2015008815A0 (en) | 2013-04-25 | 2015-10-31 | Crucell Holland Bv | Stabilized soluble prefusion rsv f polypeptides |
| MY171210A (en) | 2013-06-17 | 2019-10-02 | Janssen Vaccines & Prevention Bv | Stabilized soluble pre-fusion rsv f polypeptides |
| JP6683604B2 (ja) * | 2013-09-30 | 2020-04-22 | ヤンセン ファッシンズ アンド プリベンション ベーフェーＪａｎｓｓｅｎ Ｖａｃｃｉｎｅｓ ＆ Ｐｒｅｖｅｎｔｉｏｎ Ｂ．Ｖ． | 高密度粗製細胞培養回収物の清澄化のための方法 |
| EP3653723B1 (en) * | 2014-02-10 | 2022-07-20 | Zymo Research Corporation | Methods for nucleic acid capture |
| GB201415579D0 (en) | 2014-09-03 | 2014-10-15 | Psioxus Therapeutics Ltd | A process |
| JP6571679B2 (ja) | 2014-04-25 | 2019-09-04 | トランスレイト バイオ， インコーポレイテッド | メッセンジャーｒｎａの精製方法 |
| CA2955877A1 (en) * | 2014-07-24 | 2016-01-28 | Janssen Vaccines & Prevention B.V. | Process for the purification of poliovirus from cell cultures |
| KR20170044194A (ko) | 2014-08-27 | 2017-04-24 | 싸이오서스 테라퓨틱스 엘티디. | 아데노바이러스의 제조를 위한 공정 |
| SI3283634T1 (sl) | 2015-04-14 | 2019-08-30 | Janssen Vaccines & Prevention B.V. | Rekombinantni adenovirus, ki izraža dva transgena z dvosmernim promotorjem |
| JP7189014B2 (ja) | 2015-07-07 | 2022-12-13 | ヤンセン ファッシンズ アンド プリベンション ベーフェー | Ｒｓｖに対するワクチン |
| EP3319634B1 (en) | 2015-07-07 | 2019-08-21 | Janssen Vaccines & Prevention B.V. | Stabilized soluble pre-fusion rsv f polypeptides |
| US9663766B2 (en) | 2015-07-24 | 2017-05-30 | Bio-Rad Laboratories, Inc. | Methods for purifying adenovirus vectors |
| JP7233928B2 (ja) | 2016-04-05 | 2023-03-07 | ヤンセン ファッシンズ アンド プリベンション ベーフェー | Ｒｓｖに対するワクチン |
| DK3439672T3 (da) | 2016-04-05 | 2021-02-15 | Janssen Vaccines & Prevention Bv | Stabiliserede opløselige præfusions-rsv-f-proteiner |
| KR102573534B1 (ko) | 2016-05-12 | 2023-08-31 | 얀센 백신스 앤드 프리벤션 비.브이. | 강력하고 균형 잡힌 양방향성 프로모터 |
| ES2836598T3 (es) | 2016-05-30 | 2021-06-25 | Janssen Vaccines & Prevention Bv | Proteínas F de RSV prefusión estabilizadas |
| CN109312362B (zh) | 2016-06-20 | 2022-06-28 | 扬森疫苗与预防公司 | 有效和平衡的双向启动子 |
| GB201612248D0 (en) | 2016-07-14 | 2016-08-31 | Puridify Ltd | New process |
| JP7229151B2 (ja) | 2016-07-14 | 2023-02-27 | ヤンセン ファッシンズ アンド プリベンション ベーフェー | Ｈｐｖワクチン |
| AU2018217935B2 (en) | 2017-02-09 | 2020-04-30 | Janssen Vaccines & Prevention B.V. | Potent and short promoter for expression of heterologous genes |
| JP2020519663A (ja) | 2017-05-17 | 2020-07-02 | ヤンセン ファッシンズ アンド プリベンション ベーフェーＪａｎｓｓｅｎ Ｖａｃｃｉｎｅｓ ＆ Ｐｒｅｖｅｎｔｉｏｎ Ｂ．Ｖ． | Ｒｓｖ感染に対する防御免疫を誘導するための方法及び組成物 |
| SG11202001458SA (en) | 2017-09-15 | 2020-03-30 | Janssen Vaccines & Prevention Bv | Method for the safe induction of immunity against rsv |
| US12060583B2 (en) | 2018-04-24 | 2024-08-13 | Merck Sharp & Dohme Llc | Scalable chromatography process for purification of human cytomegalovirus |
| CN118421617A (zh) | 2018-08-24 | 2024-08-02 | 川斯勒佰尔公司 | 用于纯化信使rna的方法 |
| CN111454914A (zh) * | 2019-01-18 | 2020-07-28 | 嘉兴安宇生物科技有限公司 | 一种腺病毒快速纯化方法 |
| KR102125567B1 (ko) * | 2019-07-02 | 2020-06-22 | 한양대학교 에리카산학협력단 | 식물 엑소좀의 대량 생산 방법 |
| AU2020400034A1 (en) * | 2019-12-10 | 2022-05-26 | Repligen Corporation | Methods of preparing viral vectors |
| IL303912A (en) * | 2020-12-21 | 2023-08-01 | Pfizer | Methods and preparations for preventing excess sedimentation of nucleic acids |
| JP2023141423A (ja) * | 2022-03-24 | 2023-10-05 | 国立大学法人 東京大学 | ウイルスの精製方法 |
| CN115073549B (zh) * | 2022-07-07 | 2023-10-20 | 澳斯康生物(南通)股份有限公司 | Hek293细胞裂解液的纯化方法 |
| CN117551624A (zh) * | 2024-01-11 | 2024-02-13 | 深圳源兴基因技术有限公司 | 一种基于无血清悬浮扩增的高效回收腺病毒的方法 |
Family Cites Families (36)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2705686B1 (fr) | 1993-05-28 | 1995-08-18 | Transgene Sa | Nouveaux adénovirus défectifs et lignées de complémentation correspondantes. |
| US5851806A (en) | 1994-06-10 | 1998-12-22 | Genvec, Inc. | Complementary adenoviral systems and cell lines |
| CA2192442C (en) | 1994-06-10 | 2007-09-25 | Imre Kovesdi | Complementary adenoviral vector systems and cell lines |
| US5846782A (en) | 1995-11-28 | 1998-12-08 | Genvec, Inc. | Targeting adenovirus with use of constrained peptide motifs |
| US5559099A (en) | 1994-09-08 | 1996-09-24 | Genvec, Inc. | Penton base protein and methods of using same |
| US5965541A (en) | 1995-11-28 | 1999-10-12 | Genvec, Inc. | Vectors and methods for gene transfer to cells |
| US5770442A (en) | 1995-02-21 | 1998-06-23 | Cornell Research Foundation, Inc. | Chimeric adenoviral fiber protein and methods of using same |
| IL122614A0 (en) | 1995-06-15 | 1998-08-16 | Introgene Bv | Packaging systems for human recombinant adenovirus to be used in gene therapy |
| US6143548A (en) | 1995-08-30 | 2000-11-07 | Genzyme Corporation | Chromatographic purification of adeno-associated virus (AAV) |
| US5837511A (en) | 1995-10-02 | 1998-11-17 | Cornell Research Foundation, Inc. | Non-group C adenoviral vectors |
| CA2177085C (en) | 1996-04-26 | 2007-08-14 | National Research Council Of Canada | Adenovirus e1-complementing cell lines |
| DE19625666C1 (de) | 1996-06-26 | 1998-01-15 | Siemens Ag | Ausleseschaftung und kapazitiv messender Senser |
| AU3447097A (en) | 1996-07-01 | 1998-01-21 | Rhone-Poulenc Rorer S.A. | Method for producing recombinant adenovirus |
| DE69737107T2 (de) | 1996-11-20 | 2007-07-12 | Introgen Therapeutics Inc., Austin | Ein verbessertes verfahren zur produktion und reinigung von adenoviralen vektoren |
| JP2000509614A (ja) | 1997-03-04 | 2000-08-02 | バクスター インターナショナル インコーポレイテッド | アデノウイルスｅ１−相補性細胞系 |
| US6020191A (en) | 1997-04-14 | 2000-02-01 | Genzyme Corporation | Adenoviral vectors capable of facilitating increased persistence of transgene expression |
| US5981225A (en) | 1998-04-16 | 1999-11-09 | Baylor College Of Medicine | Gene transfer vector, recombinant adenovirus particles containing the same, method for producing the same and method of use of the same |
| US6113913A (en) | 1998-06-26 | 2000-09-05 | Genvec, Inc. | Recombinant adenovirus |
| US20030017138A1 (en) | 1998-07-08 | 2003-01-23 | Menzo Havenga | Chimeric adenoviruses |
| JP2003523169A (ja) * | 1998-12-31 | 2003-08-05 | アバンテイス・フアルマ・エス・アー | ウイルス粒子の分離法 |
| US6492169B1 (en) | 1999-05-18 | 2002-12-10 | Crucell Holland, B.V. | Complementing cell lines |
| DE19955558C2 (de) | 1999-11-18 | 2003-03-20 | Stefan Kochanek | Permanente Amniozyten-Zelllinie, ihre Herstellung und Verwendung zur Herstellung von Gentransfervektoren |
| US6544424B1 (en) | 1999-12-03 | 2003-04-08 | Refined Technology Company | Fluid filtration system |
| US20020064860A1 (en) | 2000-11-29 | 2002-05-30 | Schering Corporation | Method for purifying adenoviruses |
| ES2335657T3 (es) | 2002-04-25 | 2010-03-31 | Crucell Holland B.V. | Medios y metodos para la produccion de vectores de adenovirus. |
| DE60335672D1 (de) | 2002-05-14 | 2011-02-17 | Merck Sharp & Dohme | Verfahren zur reinigung von adenovirus |
| EP1371723A1 (en) | 2002-06-12 | 2003-12-17 | Procorde GmbH | Process for preparing an adenovirus-containing preparation |
| DK1623023T3 (da) | 2003-05-09 | 2009-03-02 | Crucell Holland Bv | Kulturer af E1-immortaliserede celler og fremgangsmåder til dyrkning deraf til forögelse af produktudbytter derfra |
| ES2329607T3 (es) | 2004-02-23 | 2009-11-27 | Crucell Holland B.V. | Metodos de purificacion de virus. |
| JP2007525984A (ja) | 2004-03-05 | 2007-09-13 | ディーエスエム アイピー アセッツ ビー．ブイ． | 連続的灌流および交互接線流による細胞培養の方法 |
| AU2005305347A1 (en) | 2004-11-03 | 2006-05-18 | Introgen Therapeutics Inc. | Method of producing and purifying of adenoviral vectors |
| ATE412737T1 (de) | 2005-04-11 | 2008-11-15 | Crucell Holland Bv | Virusreinigung mit ultrafiltration |
| SI2041259T1 (sl) | 2006-07-14 | 2016-04-29 | Dpx Holdings B.V. | Izboljšan postopek kultiviranja celic |
| CN103952376A (zh) * | 2008-09-24 | 2014-07-30 | 米迪缪尼有限公司 | 培养细胞、增殖和纯化病毒的方法 |
| PL2350268T3 (pl) | 2008-11-03 | 2015-05-29 | Crucell Holland Bv | Sposób wytwarzania wektorów adenowirusowych |
| WO2011045378A1 (en) | 2009-10-15 | 2011-04-21 | Crucell Holland B.V. | Method for the purification of adenovirus particles |
-
2010
- 2010-10-14 ES ES10771402.4T patent/ES2445713T3/es active Active
- 2010-10-14 CA CA2777116A patent/CA2777116C/en active Active
- 2010-10-14 MX MX2012004221A patent/MX2012004221A/es active IP Right Grant
- 2010-10-14 DK DK10771402.4T patent/DK2488635T3/en active
- 2010-10-14 KR KR1020127012427A patent/KR101805938B1/ko active IP Right Grant
- 2010-10-14 US US13/501,731 patent/US8470585B2/en active Active
- 2010-10-14 EP EP10771402.4A patent/EP2488635B1/en active Active
- 2010-10-14 EA EA201270557A patent/EA028875B1/ru not_active IP Right Cessation
- 2010-10-14 AU AU2010305768A patent/AU2010305768B2/en not_active Ceased
- 2010-10-14 PL PL10771402T patent/PL2488635T3/pl unknown
- 2010-10-14 CN CN201080046353.5A patent/CN102575233B/zh active Active
- 2010-10-14 WO PCT/EP2010/065436 patent/WO2011045381A1/en active Application Filing
- 2010-10-14 JP JP2012533631A patent/JP5465331B2/ja active Active
- 2010-10-14 BR BR112012008507A patent/BR112012008507B8/pt not_active IP Right Cessation
-
2012
- 2012-04-05 ZA ZA2012/02524A patent/ZA201202524B/en unknown
-
2013
- 2013-02-20 HK HK13102162.3A patent/HK1174949A1/xx not_active IP Right Cessation
Also Published As
| Publication number | Publication date |
|---|---|
| ZA201202524B (en) | 2012-12-27 |
| ES2445713T3 (es) | 2014-03-04 |
| BR112012008507B8 (pt) | 2021-05-25 |
| KR20120093957A (ko) | 2012-08-23 |
| WO2011045381A1 (en) | 2011-04-21 |
| AU2010305768B2 (en) | 2015-05-14 |
| CN102575233A (zh) | 2012-07-11 |
| HK1174949A1 (en) | 2013-06-21 |
| EA201270557A1 (ru) | 2012-09-28 |
| BR112012008507B1 (pt) | 2021-05-04 |
| AU2010305768A1 (en) | 2012-04-12 |
| BR112012008507A2 (pt) | 2015-09-15 |
| US20120202268A1 (en) | 2012-08-09 |
| CA2777116C (en) | 2020-09-01 |
| US8470585B2 (en) | 2013-06-25 |
| EA028875B1 (ru) | 2018-01-31 |
| EP2488635B1 (en) | 2013-11-20 |
| EP2488635A1 (en) | 2012-08-22 |
| JP2013507141A (ja) | 2013-03-04 |
| JP5465331B2 (ja) | 2014-04-09 |
| KR101805938B1 (ko) | 2018-01-10 |
| DK2488635T3 (en) | 2014-02-24 |
| CN102575233B (zh) | 2014-07-16 |
| MX2012004221A (es) | 2012-06-08 |
| CA2777116A1 (en) | 2011-04-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| HK1174949A1 (en) | Process for adenovirus purification from high cell density cultures | |
| HK1174948A1 (en) | Method for the purification of adenovirus particles from high cell density cultures | |
| GB0914195D0 (en) | Vessel for culturing cells | |
| SI2384308T1 (sl) | Vodikov postopek | |
| EP2411501A4 (en) | CELL OR TISSUE CULTURE APPARATUS | |
| EP2257986A4 (en) | PROCESS FOR MANUFACTURING SOLAR CELLS | |
| EP2500418A4 (en) | NEW METHOD FOR THE PRODUCTION OF DIFFERENTIATED CELLS | |
| EP2443225A4 (en) | BIOREACTOR SYSTEM | |
| HK1243731A1 (zh) | 用於培養貼壁細胞的方法 | |
| EP2510020A4 (en) | MULTI-REACTOR PROCESS | |
| ZA201205174B (en) | Anaerobic reactor | |
| EP2506273A4 (en) | Cooling structure for magnet-fitted reactor | |
| EP2419192A4 (en) | APPARATUS FOR PURIFYING WATER | |
| GB0800524D0 (en) | Cell culture system | |
| PL2308969T3 (pl) | Sposób przyspieszania namnażania komórek | |
| EP2406369A4 (en) | METHOD FOR MONITORING CELLULAR CULTURE | |
| GB0817990D0 (en) | Cell culture vessel | |
| GB0913469D0 (en) | Cell support | |
| EP2414062A4 (en) | DEVICE FOR CLEANING WATER | |
| EP2511364A4 (en) | METHOD FOR CULTURING CELLS OR MICRO-ORGANISMS | |
| EP2271739A4 (en) | CELL CULTURE PRODUCT | |
| ZA201200628B (en) | Fermentation process | |
| GB2475401B (en) | Improved agitated cell reactor | |
| HUE045635T2 (hu) | Eljárás továbbfejlesztett egysejtes klónozásra | |
| GB0906424D0 (en) | Use of compounds for differentiation of cells |