ES2539105T3 - Células mesenquimatosas y osteoblastos procedentes de células madre embrionarias humanas - Google Patents

Células mesenquimatosas y osteoblastos procedentes de células madre embrionarias humanas Download PDF

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ES2539105T3
ES2539105T3 ES02756367.5T ES02756367T ES2539105T3 ES 2539105 T3 ES2539105 T3 ES 2539105T3 ES 02756367 T ES02756367 T ES 02756367T ES 2539105 T3 ES2539105 T3 ES 2539105T3
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cells
embryonic stem
human embryonic
mesenchymal
osteoblast
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Chunhui Xu
R. Scott Thies
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Asterias Biotherapeutics Inc
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    • C12N5/0602—Vertebrate cells
    • C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
    • C12N5/0654—Osteocytes, Osteoblasts, Odontocytes; Bones, Teeth
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Abstract

Un medio de cultivo para obtener osteoprogenitores y/u osteoblastos a partir de células madre embrionarias humanas (hES) o su progenie, que comprende BMP4, dexametasona y ácido ascórbico o uno de sus análogos.

Description

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E02756367
10-06-2015
Se transdujo una subpoblación para expresar la transcriptasa inversa de la telomerasa humana (hTERT). Esto se logró infectando con un constructo retrovírico pBABE puro hTERT, que contenía la secuencia codificante de hTERT controlada por MoLV LTR y el gen de resistencia a la puromicina controlado por el promotor temprano SV40. Se reemplazó el medio de crecimiento con una mezcla que contenía 5 mL de patrón retrovírico (1 x 106 pfu/mL) y 4 µg/mL de polybrene y se incubó a 37 ºC. Después de 8 h, se añadieron 5 mL adicionales de la mezcla retrovirus/polybrene y las células se incubaron a 37 ºC. Al día siguiente, la mezcla retrovirus/polybrene se retiró y se reemplazó con medio de crecimiento fresco. Al día siguiente se reemplazó el medio con medio de crecimiento suplementado con 0.5 microgramos/mL de puromicina. Se dividieron las células una vez a la semana con una relación de 1:4 durante 8 semanas en medio que contenía puromicina y se probaron para determinar su actividad telomerasa.
La Figura 2 (Cuadro A) muestra la morfología de la línea celular telomerizada HEF1. El cuadro B (debajo) muestra la actividad telomerasa, según se ha medido con el ensayo TRAP. Las células transducidas con el casete de expresión de hTERT mostraron una actividad telomerasa positiva 20 o 65 días después de la transducción. La línea celular sin transducir, o las células transducidas con el vector de control no mostraron actividad telomerasa. Tanto las células HEF1 transducidas con hTERT como las células transducidas con el vector de control, duplicaron su cantidad aproximadamente una vez cada 2 días, hasta el día 38, cuando las células de control cesaron de dividirse. Las células transfectadas con hTERT continuaron proliferando más allá de los 60 días (30 duplicaciones) con una velocidad de crecimiento constante.
El medio de crecimiento de las células ES se acondicionó como en el Ejemplo 8, utilizando células HEF1 irradiadas con 6000 rad, y se sembraron con una densidad de ~4.1 a 5.5 x 104 células cm-2. Se probó el medio para determinar su capacidad de favorecer el crecimiento de la línea celular hES H9 cultivada en un sustrato Matrigel®. Las células hES se mantuvieron utilizando el medio acondicionado para HEF1 durante más de 4 pases, mostraron morfología de células ES indiferenciadas y mantuvieron la expresión de hTERT y Oct-4.
Ejemplo de referencia 3: Diferenciación adicional en células similares a osteoblastos
Las células ES humanas (línea celular H1, pase 30) se mantuvieron en condiciones exentas de alimentadoras, como se ha descrito anteriormente. Para su uso en este experimento, las células hES se sembraron con una densidad de ~1 × 105 cm-2 en Matrigel® en medio acondicionado para mEF. Las células HEF1 telomerizadas se colocaron en placas con una densidad de 3.1 × 103 cm-2 en DMEM con un 10% de FBS, un 1% de aminoácidos no esenciales y Lglutamina 2 mM. Las células madre mesenquimatosas humanas (hMSC) normales se obtuvieron de BioWhittaker Inc., MD (una filial de Cambrex Co.). Se mantuvieron en medio de crecimiento para MSC (BioWhittaker Part n.º PT3001) de acuerdo con las indicaciones del proveedor. La línea celular de fibroblastos BJ5ta (Bodnar et al., Science 279:349, 1998) se mantuvo en un medio estándar constituido por un 10% de FBS en 1:3 M199/DMEM.
Dos días después del último pase, cada medio de cultivo se reemplazó por medio de inducción de osteoblastos (OIM) para inducir la diferenciación. La base del OIM fue el medio de crecimiento para MSC (ClonTech n.º de cat. PT-3238) (patente de los EE. UU. 5 486 359) suplementado con dexametasona 0.1 µM, ácido 2-fosfato ascórbico 5 µM, β-glicerofosfato 10 mM y 100 ng/mL de BMP-4. Las células se alimentaron con OIM fresco cada 2-3 días.
Después de 11 días en OIM, todas las células mostraron cambios en la morfología celular. Las células HEF1, hMSC y células BJ cambiaron de una forma en husillo a una forma cuboidal y algunas células se volvieron planas. Las células hES mostraron una morfología heterogénea que parecía ser una población diferenciada mixta.
Las células se fijaron con paraformaldehído al 2% en PBS durante 20 min, se lavaron con PBS y se analizaron para determinar los marcadores de osteoblastos. Se detectó la fosfatasa alcalina (AP) con sustrato Vector (Vector Laboratories, Inc., Burlingame, CA). La expresión de AP estuvo claramente ubicada en agrupaciones de células, células H1 diferenciadas así como también células HEF1, BJ y hMSC.
Las proteínas matriciales producidas por los osteoblastos, colágeno-1 y osteocalcina se detectaron mediante inmunotinción. Se permeabilizaron los cultivos mediante tratamiento con EtOH al 100% durante 2 min. Después de los lavados en PBS, se incubaron los cultivos con un 5% de suero de cabra normal en PBS durante 2 h y a continuación con anticuerpo de conejo primario contra el colágeno-1 (1:10, Monosan n.º de cat. P5041) u osteocalcina (1:50, Biomedical Technologies Inc. n.º de cat. 13T593). La tinción se reveló con el anticuerpo secundario anti-inmunoglobulina de conejo producido en cabra marcado con FITC (1:100, Southern Biotechnology Associates inc. n.º de cat. 4050-02).
La Figura 3 muestra los resultados. Los cuadros A y B muestran los resultados inmunocitoquímicos de los marcadores osteocalcina y colágeno-1. El cuadro C muestra la tinción de la actividad fosfatasa alcalina. Estos rasgos son característicos de las células del linaje de osteoblastos.
Estos datos son coherentes con la hipótesis de que tanto las células hES como las células HEF1 tienen la capacidad de generar osteoblastos cuando se someten a un protocolo de diferenciación apropiado in vitro.
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Claims (1)

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ES02756367.5T 2001-07-06 2002-07-03 Células mesenquimatosas y osteoblastos procedentes de células madre embrionarias humanas Expired - Lifetime ES2539105T3 (es)

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US30373201P 2001-07-06 2001-07-06
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PCT/US2002/020998 WO2003004605A2 (en) 2001-07-06 2002-07-03 Mesenchymal cells and osteoblasts from human embryonic stem cell

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Families Citing this family (38)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7410798B2 (en) 2001-01-10 2008-08-12 Geron Corporation Culture system for rapid expansion of human embryonic stem cells
JP2005503759A (ja) * 2001-01-24 2005-02-10 アメリカ合衆国 幹細胞の膵臓内分泌細胞への分化方法
US20030211605A1 (en) * 2001-05-01 2003-11-13 Lee Sang-Hun Derivation of midbrain dopaminergic neurons from embryonic stem cells
GB0222846D0 (en) 2002-10-03 2002-11-06 Choo Yen Cell culture
HUE028026T2 (en) 2002-12-16 2016-11-28 Technion Res & Dev Foundation Breeding system free of native cells, free of foreign matter for human embryonic stem cells
US20030224411A1 (en) * 2003-03-13 2003-12-04 Stanton Lawrence W. Genes that are up- or down-regulated during differentiation of human embryonic stem cells
JP2005304443A (ja) * 2004-04-26 2005-11-04 Institute Of Physical & Chemical Research 前駆間葉系幹細胞
AU2005243158B2 (en) * 2004-05-07 2010-03-25 Wisconsin Alumni Research Foundation Method of forming mesenchymal stem cells from embryonic stem cells
WO2005113749A2 (en) * 2004-05-14 2005-12-01 Becton, Dickinson And Company Stem cell populations and methods of use
GB2431165B (en) 2004-07-13 2009-04-01 Geron Corp Medium for growing human embryonic stem cells
AU2006235211B2 (en) 2005-04-12 2010-10-14 Mesoblast, Inc. Isolation of adult multipotential cells by tissue non-specific alkaline phosphatase
KR20130100221A (ko) 2005-06-22 2013-09-09 제론 코포레이션 인간 배아 줄기 세포의 현탁 배양
JP4714741B2 (ja) * 2005-07-29 2011-06-29 学校法人松本歯科大学 歯の再生方法
WO2007026353A2 (en) 2005-08-29 2007-03-08 Technion Research & Development Foundation Ltd. Media for culturing stem cells
BRPI0617084A2 (pt) * 2005-09-02 2011-07-12 Agency Science Tech & Res método, linhagem de célula progenitora, célula diferenciada e método para gerar uma célula diferenciada de uma célula-tronco (es) embrionária
GB0526664D0 (en) * 2005-11-30 2006-02-08 Plasticell Ltd Method
EP1981971A4 (en) * 2006-01-11 2009-08-12 Technion Res & Dev Foundation PROGENITORS OF CONNECTIVE TISSUES OBTAINED FROM ADULT STEM CELLS FOR TISSUE GENE
CA2640794C (en) * 2006-02-16 2015-04-14 Universite Libre De Bruxelles A method for osteogenic differentiation of bone marrow stem cells (bmsc) and uses thereof
WO2007122233A1 (en) * 2006-04-25 2007-11-01 Vrije Universiteit Brussel Preparation of mesenchymal progenitor cells, particularly osteogenic progenitor cells
ES2704401T3 (es) 2006-08-02 2019-03-18 Technion Res & Dev Foundation Métodos de expansión de células madre embrionarias en un cultivo en suspensión
JP2010500047A (ja) * 2006-08-15 2010-01-07 エージェンシー フォー サイエンス，テクノロジー アンド リサーチ 間葉系幹細胞馴化培地
US8153359B2 (en) * 2006-10-02 2012-04-10 Cellartis Ab Toxicity assay based on human blastocyst-derived stem cells and progenitor cells
KR100937456B1 (ko) * 2007-08-01 2010-01-19 한국생명공학연구원 인간배아줄기세포를 조골세포 직계열로 분화시키는 방법
WO2010105311A1 (en) * 2009-03-20 2010-09-23 Angioblast Systems, Inc. Production of reprogrammed pluripotent cells
AU2010266016B2 (en) 2009-06-25 2016-05-26 Asterias Biotherapeutics, Inc. Differentiated pluripotent stem cell progeny depleted of extraneous phenotypes
KR101135636B1 (ko) * 2009-10-27 2012-04-17 서울대학교산학협력단 인간 만능줄기세포로부터 중배엽 줄기세포를 생산하는 방법 및 이에 의해 생성된 중배엽 줄기세포
ES2779048T3 (es) 2009-11-12 2020-08-13 Technion Res & Dev Foundation Medios de cultivo, cultivos celulares y métodos de cultivo de células madre pluripotentes en un estado indiferenciado
AU2011236653B2 (en) 2010-04-08 2016-10-27 The University Court Of The University Of Edinburgh Chondrogenic progenitor cells, protocol for derivation of cells and uses thereof
WO2011124741A1 (es) * 2010-04-08 2011-10-13 Fundación Progreso Y Salud Uso de un medio de cultivo condicionado por células madre mesenquimales para la diferenciación de células madre pluripotentes humanas
EP2582793B1 (en) 2010-06-15 2017-09-06 Cellular Dynamics International, Inc. A compendium of ready-built stem cell models for interrogation of biological response
US20140329314A1 (en) 2011-03-29 2014-11-06 Christopher O'Sullivan Enriched populations of cardiomyocyte lineage cells from pluripotent stem cells
CN103083653B (zh) * 2011-10-28 2015-07-22 辽宁成大动物药业有限公司 一种采用微载体高密度细胞培养技术生产猪瘟活疫苗的方法
CN103777009B (zh) * 2012-10-18 2016-03-02 辽宁成大动物药业有限公司 一种使用辣根过氧化物酶标抗体检测猪瘟弱毒病毒滴度的方法
US20170044500A1 (en) 2014-04-24 2017-02-16 Board Of Regents, The University Of Texas System Application of induced pluripotent stem cells to generate adoptive cell therapy products
US11285177B2 (en) 2018-01-03 2022-03-29 Globus Medical, Inc. Allografts containing viable cells and methods thereof
CN108570448B (zh) * 2018-01-26 2019-04-02 皓昇莱生物制药有限公司 一种高效的hPSCs向MSCs分化的方法
CN114127266A (zh) * 2019-04-23 2022-03-01 雪拉托兹治疗株式会社 肌肉骨骼系统干细胞的选择性分化调节方法
CN113462642A (zh) * 2021-08-12 2021-10-01 呈诺再生医学科技(珠海横琴新区)有限公司 间充质干细胞的快速诱导分化方法、试剂盒及其应用

Family Cites Families (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5486359A (en) * 1990-11-16 1996-01-23 Osiris Therapeutics, Inc. Human mesenchymal stem cells
US5654186A (en) * 1993-02-26 1997-08-05 The Picower Institute For Medical Research Blood-borne mesenchymal cells
US5681701A (en) * 1993-07-12 1997-10-28 Mayo Foundation For Medical Education And Research Immortalized human fetal osteoblastic cells
US5691175A (en) * 1993-07-12 1997-11-25 Mayo Foundation For Medical Education And Research Immortalized human fetal osteoblastic cells
US5972703A (en) * 1994-08-12 1999-10-26 The Regents Of The University Of Michigan Bone precursor cells: compositions and methods
US5843780A (en) * 1995-01-20 1998-12-01 Wisconsin Alumni Research Foundation Primate embryonic stem cells
GB9516268D0 (en) * 1995-08-08 1995-10-11 Danbiosyst Uk Compositiion for enhanced uptake of polar drugs from the colon
ATE288480T1 (de) * 1995-11-16 2005-02-15 Univ Case Western Reserve Chondrogene in vitro induktion von menschlichen mensenchymalen stammzellen
US6482231B1 (en) * 1995-11-20 2002-11-19 Giovanni Abatangelo Biological material for the repair of connective tissue defects comprising mesenchymal stem cells and hyaluronic acid derivative
US6200606B1 (en) * 1996-01-16 2001-03-13 Depuy Orthopaedics, Inc. Isolation of precursor cells from hematopoietic and nonhematopoietic tissues and their use in vivo bone and cartilage regeneration
US6090622A (en) * 1997-03-31 2000-07-18 The Johns Hopkins School Of Medicine Human embryonic pluripotent germ cells
US6077987A (en) * 1997-09-04 2000-06-20 North Shore-Long Island Jewish Research Institute Genetic engineering of cells to enhance healing and tissue regeneration
AU1197699A (en) * 1997-10-23 1999-05-10 Geron Corporation Methods and materials for the growth of primate-derived primordial stem cells
US6667176B1 (en) 2000-01-11 2003-12-23 Geron Corporation cDNA libraries reflecting gene expression during growth and differentiation of human pluripotent stem cells
CA2349415A1 (en) * 1998-11-09 2000-05-18 Monash University Embryonic stem cells
NZ516738A (en) * 1999-07-20 2004-01-30 Univ Southern California Identification of pluripotent pre-mesenchymal, pre-hematopoietic progenitor cells expressing a unique molecular marker
NZ518191A (en) * 1999-10-15 2004-01-30 Advanced Cell Tech Inc Methods of producing differentiated progenitor cells and lineage-defective embryonic stem cells
AU1618201A (en) * 1999-11-19 2001-05-30 Children's Medical Center Corporation Methods for inducing chondrogenesis and producing de novo cartilage in vitro
GB0003930D0 (en) * 2000-02-18 2000-04-12 King S College London Cell
US20040224403A1 (en) * 2001-12-07 2004-11-11 Robarts Research Institute Reconstituting hematopoietic cell function using human embryonic stem cells

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