ES2525718T3 - Reparación y regeneración de tejido renal mediante células derivadas de tejido de cordón umbilical humano - Google Patents

Reparación y regeneración de tejido renal mediante células derivadas de tejido de cordón umbilical humano Download PDF

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ES2525718T3
ES2525718T3 ES08835175.4T ES08835175T ES2525718T3 ES 2525718 T3 ES2525718 T3 ES 2525718T3 ES 08835175 T ES08835175 T ES 08835175T ES 2525718 T3 ES2525718 T3 ES 2525718T3
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umbilical cord
protein
tissue
cord tissue
human umbilical
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David Colter
Anna Gosiewska
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DePuy Synthes Products Inc
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/44—Vessels; Vascular smooth muscle cells; Endothelial cells; Endothelial progenitor cells
    • A—HUMAN NECESSITIES
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    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0603—Embryonic cells ; Embryoid bodies
    • C12N5/0605—Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells

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Abstract

Células obtenidas de tejido de cordón umbilical humano para su uso en el tratamiento de un paciente con lesión o insuficiencia renal aguda o enfermedad renal crónica, en el que el tejido de cordón umbilical está sustancialmente libre de sangre, y en el que las células son capaces de autorrenovarse y desarrollarse en cultivo y tienen el potencial de diferenciarse; necesitan L-valina para crecer; son capaces de crecer en atmósferas que contienen oxígeno del 5% al 20%; no producen CD117 ni HLA-DR; expresan alfa-actina de músculo liso; y expresan, en comparación con una célula madre mesenquimal, célula de médula ósea de cresta ilíaca o fibroblasto humano, mayores niveles de receptor 1 de lipoproteínas de baja densidad oxidadas, interleucina 8 o reticulón 1.

Description

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E08835175
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Tabla 4
5
15
25
35
45
55
Los genes demostraron tener una expresión disminuida en las células derivadas de tejido de cordón umbilical en comparación con otras líneas celulares ensayadas
Genes disminuidos en las células derivadas de placenta y derivadas de tejido de cordón umbilical
ID del conjunto de sondas
Nombre el gen Número de registro del NCBI
210135_ s_at
caja homeótica de la baja estatura 2 AF022654.1
205824_at
proteína 2 de choque térmico de 27 kDa NM_001541.1
209687_at
ligando 12 de quimiocina (motivo C-X-C ) (factor 1 derivado de células del estroma) U19495.1
203666_at
ligando 12 de quimiocina (motivo C-X-C ) (factor 1 derivado de células del estroma) NM_000609.1
212670_at
elastina (estenosis aórtica supravalvular, síndrome de Williams-Beuren) AA479278
213381_at
ARNm de Homo sapiens; ADNc DKFZp586M2022 (del clon DKFZp586M2022) N91149
206201_s_at
caja homeótica de mesénquima 2 (caja homeótica específica de la detención del crecimiento) NM_005924.1
205817_at
homólogo 1 de caja homeótica de “sine oculis” (Drosophila) NM_005982.1
209283_at
cristalino, alfa B AF007162.1
212793_at
activador 2 de la morfogénesis asociado a “disheveled” BF513244
213488_at
proteína DKFZP586B2420 AL050143.1
209763_at
similar a neuralina 1 AL049176
205200_at
tetranectina (proteína de unión a plasminógeno) NM_003278.1
205743_at
de homología con src tres (SH3) y dominio rico en cisteína NM_003149.1
200921_s_at
gen 1 de translocación de linfocitos B, anti-proliferativa NM_001731.1
206932_at
colesterol 25-hidroxilasa NM_003956.1
204198_s_at
factor de transcripción 3 relacionado con enanismo AA541630
219747_at
proteína hipotética FLJ23191 NM_024574.1
204773_at
receptor de interleucina 11, alfa NM_004512.1
202465_at
potenciador de procolágeno C-endopeptidasa NM_002593.2
203706_s_at
homólogo 7 de “frizzled“ (Drosophila) NM_003507.1
212736_at
gen hipotético BC008967 BE299456
214587_at
colágeno, tipo VIII, alfa 1 BE877796
201645_at
tenascina C (hexabraquion) NM_002160.1
210239_at
proteína 5 de la caja homeótica de “iroquois“ U90304.1
203903_s_at
hepaestina NM_014799.1
205816_at
integrina, beta 8 NM_002214.1
203069_at
glicoproteína 2 de las vesículas sinápticas NM_014849.1
213909_at
ADNc FLJ12280 fis de Homo sapiens, clon MAMMA1001744 AU147799
206315_at
factor 1 similar a receptor de citocinas NM_004750.1
204401_at
canal de potasio activado por calcio de conductancia intermedia/baja, subfamilia N, miembro 4 NM_002250.1
216331_at
integrina, alfa 7 AK022548.1
209663_s_at
integrina, alfa 7 AF072132.1
213125_at
proteína DKFZP586L151 AW007573
202133_at
coactivador de la transcripción con motivo de unión a PDZ (TAZ) AA081084
206511_s_at
homólogo 2 de la caja homeótica de “sine oculis” (Drosophila) NM_016932.1
213435_at
proteína KIAA1034 AB028957.1
206115_at
de respuesta de crecimiento precoz 3 NM_004430.1
213707_s_at
caja homeótica 5 de “distal-less” NM_005221.3
19
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218181_s_at
proteína hipotética FLJ20373 NM_017792.1
209160_ at
familia 1 de aldo-ceto reductasa, miembro C3 (3-alfa hidroxiesteroide deshidrogenasa, tipo II) AB018580.1
213905_x_at
biglicano AA845258
201261_x_at
biglicano BC002416.1
202132_at
coactivador de la transcripción con motivo de unión a PDZ (TAZ) AA081084
214701_s_at
fibronectina 1 AJ276395.1
213791_at
proencefalina NM_006211.1
205422_s_at
integrina, similar a beta 1 (con dominios de repetición de tipo EGF) NM_004791.1
214927_at
clon EUROIMAGE 1968422 de ADNc de inserto de longitud completa de ARNm de Homo sapiens AL359052.1
206070_s_at
EphA3 AF213459.1
212805_at
proteína KIAA0367 AB002365.1
219789_at
receptor C del péptido natriurético/guanilato ciclasa C (receptor C del péptido atrial natriurético) AI628360
219054_at
proteína hipotética FLJ14054 NM_024563.1
213429_at
ARNm de Homo sapiens; ADNc DKFZp564B222 (del clon DKFZp564B222) AW025579
204929_s_at
proteína 5 de membrana asociada a vesículas (miobrevina) NM_006634.1
201843_s_at
proteína 1 de la matriz extracelular de tipo fibulina que contienen EGF NM_004105.2
221478_at
similar a proteína 3 de interacción con BCL2/adenovirus E1B 19kDa AL132665.1
201792_at
proteína 1 de unión a AE NM_001129.2
204570_at
polipéptido 1 de la subunidad VIIa de la citocromo c oxidasa (músculo) NM_001864.1
201621_at
neuroblastoma, supresión de tumorigenicidad 1 NM_005380.1
202718_at
proteína 2 de unión al factor de crecimiento insulinoide, de 36kDa NM_000597.1
Las tablas 5, 6 y 7 muestran la expresión de genes aumentada en fibroblastos humanos (Tabla 5), células ICBM (Tabla 6) y MSC (Tabla 7).
Tabla 5
45
55
Genes que demostraron tener una expresión aumentada en los fibroblastos en comparación con las demás líneas celulares ensayadas
Genes aumentados en fibroblastos
fosfatasa de doble especificidad 2
proteína KIAA0527
ADNc de Homo sapiens: FLJ23224 fis, clon ADSU02206
dineína, citoplasmática, polipéptido intermedio 1
anquirina 3, nódulo de Ranvier (anquirina G)
inhibina, beta A (activina A, activina AB, polipéptido alfa)
ectonucleótido pirofosfatasa/fosfodiesterasa 4 (función supuesta)
proteína KIAA1053
proteína asociada a microtúbulos 1A
proteína con dedos de zinc 41
proteína HSPC019
ADNc de Homo sapiens: FLJ23564 fis, clon LNG10773
ARNm de Homo sapiens; ADNc DKFZp564A072 (del clon DKFZp564A072)
Proteína LIM (similar a enigma de unión a proteína quinasa C de rata)
inhibidor del potenciador del gen del polipéptido de cadena ligera kappa en los linfocitos B, proteína asociada a complejo quinasa
proteína hipotética FLJ22004
Secuencia de ARNm humano (clon CTG-A4)
EST, Moderadamente similares al factor 2 de tipo receptor de citocinas; precursor del receptor de citocinas CRL2 [Homo sapiens]
20
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5
15
25
35
45
55
IM042479 Medida inicial de la proliferación del receptor Control de autoestimulación (células autólogas tratadas con mitomicina C) Donante alogénico MLR IM04-2477 (tratado con mitomicina C) MLR con línea celular (célula tratada con mitomicina C, tipo A) SI (donante) SI (línea celular) IM042480 Medida inicial de la proliferación del receptor Control de autoestimulación (células autólogas tratadas con mitomicina C) Donante alogénico MLR IM04-2477 (tratado con mitomicina C) MLR con línea celular (célula tratada con mitomicina C, tipo A) SI (donante) SI (línea celular) IM042481 Medida inicial de la proliferación del receptor Control de autoestimulación (células autólogas tratadas con mitomicina C) Donante alogénico MLR IM04-2477 (tratado con mitomicina C) MLR con línea celular (célula tratada con mitomicina C, tipo A) SI (donante) SI (línea celular) Número de análisis Sistema de cultivo IM042482 Medida inicial de la proliferación del receptor Control de autoestimulación (células autólogas tratadas con mitomicina C) Donante alogénico MLR IM04-2477 (tratado con mitomicina C) MLR con línea celular (célula tratada con mitomicina C, tipo A) SI (donante) SI (línea celular)
530 508 701 567 25593 24732 5086 3932 1192 854 2963 993 25416 29721 2596 5076 695 451 738 1252 13177 24885 4495 3671 ID de placa: Placa 2 Repetic iones 1 2 432 533 1459 633 24286 30823 2762 1502 527 1111 22707 1497 1330 2197 23757 3426 555 464 15444 4674 3 274 598 31346 6723 521,7 793,0 24344,0 3505,0 47 7 1125,3 2051,0 26298,0 3699,3 23 3 567,0 818,0 17835,3 4280,0 31 8 Media 413.0 896.7 28818.3 3662.3 70 9 11,93 283,43 1481,61 1832,21 244,90 993,08 3078,27 1262,39 122,44 400,04 6209,52 534,95 DE 130.54 487.31 3933.82 2724.46 2,3 35,7 6,1 52,3 21,8 48,4 11,7 34,1 21,6 48,9 34,8 12,5 CV 31.6 54.3 13.7 74.4
27
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E08835175
10-12-2014
17,45
25,46 269,38
18,56
78,26
18,41
17,30
Promedio
17,33 54,25 97,28 114,4 3
Est:
3,04 36,41 64,75 94,15
EEM
0,81 10,51 19,52 54,36
La Tabla 20 y la Figura 4 muestran las mediciones de SCr de las muestras de suero del día 5. El tratamiento con HBSS y mMSC dio como resultado valores medios de creatinina de 0,74 mg/dl y 0,66 mg/dl, respectivamente. Se observó una reducción estadísticamente significativa en la SCr después del tratamiento con hMSC o hUTC a menor dosis (P < 0,03). El tratamiento con hMSC dio como resultado un valor medio de SCr de 0,47 mg/dl y el tratamiento con hUTC a menor dosis dio como resultado un valor medio de SCr de 0,57 mg/dl.
Tabla 20
25
35
45
Creatinina sérica. La Crs se midió el día 5 después del trasplante de las células. Desviación estándar (est.). Error estándar de la media (EEM)
Grupo de tratamiento
Material de ensayo Creatinina mg/dl Media Est. EEM Valor de p
1
HBSS HBSS HBSS HBSS HBSS 0,64 0,72 0,72 0,89 0,73 0,74 0,09 0,04 na
2
mMSC mMSC mMSC mMSC mMSC 0,64 0,72 0,72 0,48 0,72 0,66 0,10 0,05 0,213
3
hMSC hMSC hMSC hMSC 0,24 0,27 0,56 0,56 0,47 0,21 0,09 0,008
4
hUTC hUTC hUTC hUTC 0,64 0,45 0,56 0,64 0,57 0,08 0,03 0,028
5
hUTC hUTC hUTC hUTC 1,28 0,26 0,48 0,72 0,68 0,38 0,17 0,789
El día 7 después del trasplante de las células, se sacrificaron dos ratones por grupo de tratamiento y se fijaron los tejidos mediante perfusión transcardíaca con solución salina, seguido de perfusión e inmersión en paraformaldehído al 4%. Se extirparon los riñones de los ratones y se procesaron para la histología. A continuación se evaluaron los cortes teñidos con H&E para la lesión histológica. Dos patólogos entrenados realizaron
55 evaluaciones con enmascaramiento y puntuaron el grado de degeneración tubular renal.
Se midió cualitativamente la necrosis tubular en los cortes histológicos de animales tratados con HBSS, hMSC y hUTC a menor dosis. Se puntuó la degeneración tubular en una escala que iba de 1 a 4 (1 = daño mínimo, 2 = daño leve, 3 = daño moderado, 4 = daño grave). Como se muestra en la Tabla 21 y en la Figura 5, el tratamiento con vehículo y con hMSC dio como resultado una degeneración tubular leve, con una puntuación media de lesión de 2,5. Sin embargo, un tratamiento con hUTC a menor dosis dio como resultado daño tubular mínimo con una puntuación media de lesión de 1,9, que indicaba el potencial renoprotector de las hUTC.
65
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Claims (1)

  1. imagen1
    imagen2
ES08835175.4T 2007-10-05 2008-10-03 Reparación y regeneración de tejido renal mediante células derivadas de tejido de cordón umbilical humano Active ES2525718T3 (es)

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US977775P 2007-10-05
PCT/US2008/078800 WO2009046335A1 (en) 2007-10-05 2008-10-03 Repair and regeneration of renal tissue using human umbilical cord tissue-derived cells

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