RESUMEN
BACKGROUND AIMS: Human embryonic stem (hES) cells hold great potential for cell therapy and regenerative medicine because of their pluripotency and capacity for self-renewal. The conditions used to derive and culture hES cells vary between and within laboratories depending on the desired use of the cells. Until recently, stem cell culture has been carried out using feeder cells, and culture media, that contain animal products. Recent advances in technology have opened up the possibility of both xeno-free and feeder-free culture of stem cells, essential conditions for the use of stem cells for clinical purposes. To date, however, there has been limited success in achieving this aim. METHODS, RESULTS AND CONCLUSIONS: Protocols were developed for the successful derivation of two normal and three specific mutation-carrying (SMC) (Huntington's disease and myotonic dystrophy 1) genomically stable hES cell lines, and their adaptation to feeder-free culture, all under xeno-free conditions.
Asunto(s)
Diferenciación Celular , Técnicas de Cocultivo , Células Madre Embrionarias/fisiología , Animales , Supervivencia Celular , Células Cultivadas , Medio de Cultivo Libre de Suero , Células Madre Embrionarias/efectos de los fármacos , Células Madre Embrionarias/trasplante , Células Nutrientes/citología , Células Nutrientes/fisiología , Humanos , Enfermedad de Huntington/genética , Enfermedad de Huntington/terapia , Distrofia Miotónica/genética , Distrofia Miotónica/terapiaRESUMEN
Human embryonic stem cell (ESC) research has attracted wide media coverage. It has been headline news for the past several months, revealing the complex case of Professor Hwang Woo-Suk and the scientific fraud where he purported to have created the first human patient-specific stem cell lines generated by cell nucleus replacement (CNR). To ethically obtain the raw materials (eggs, sperm and embryos) for human ESC research is an enormous challenge, yet essential if this research is to proceed in its quest to try to deliver some of the expectations placed upon it: developing treatments and possible cures for a range of serious diseases. This article examines some of the ethical issues surrounding human ESC research using the four principles frequently applied to healthcare and medical research; autonomy, beneficence, non-maleficence and justice. The author strives to ask questions throughout which will encourage debate and discussion.
Asunto(s)
Técnicas de Cultivo de Embriones/ética , Ética en Investigación , Creación de Embriones para Investigación/ética , Trasplante de Células Madre/ética , Actitud Frente a la Salud , Criopreservación/ética , Destinación del Embrión/ética , Fertilización In Vitro/ética , Educación en Salud , Conocimientos, Actitudes y Práctica en Salud , Humanos , Diagnóstico Preimplantación/ética , Ética Basada en Principios , Mala Conducta Científica/ética , Cambio Social , Reino UnidoRESUMEN
The KCL032 human embryonic stem cell line was derived from a normal healthy blastocyst donated for research. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment and under current Good Manufacturing Practice (cGMP) standards. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Asunto(s)
Técnicas de Cultivo de Célula/métodos , Línea Celular/citología , Células Madre Embrionarias Humanas/citología , Biomarcadores/metabolismo , Diferenciación Celular , HumanosRESUMEN
The KCL016 human embryonic stem cell line was derived from an embryo donated for research that carried an autosomal dominant mutation affecting splicing site of the VHL gene encoding von Hippel-Lindau tumor suppressor E3 ubiquitin protein ligase (676+3A>T). The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Asunto(s)
Técnicas de Cultivo de Célula/métodos , Línea Celular/citología , Células Madre Embrionarias Humanas/citología , Mutación/genética , Proteína Supresora de Tumores del Síndrome de Von Hippel-Lindau/genética , Biomarcadores/metabolismo , Diferenciación Celular , Femenino , Humanos , Masculino , LinajeRESUMEN
The KCL038 human embryonic stem cell line was derived from a normal healthy blastocyst donated for research. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment and under current Good Manufacturing Practice (cGMP) standards. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Asunto(s)
Células Madre Embrionarias Humanas/citología , Fosfatasa Alcalina/metabolismo , Blastocisto/citología , Línea Celular , Supervivencia Celular , Hibridación Genómica Comparativa , Genotipo , Prueba de Histocompatibilidad , Células Madre Embrionarias Humanas/metabolismo , Humanos , Masculino , Microscopía Fluorescente , Persona de Mediana EdadRESUMEN
The KCL037 human embryonic stem cell line was derived from a normal healthy blastocyst donated for research. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment and under current Good Manufacturing Practice (cGMP) standards. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Asunto(s)
Células Madre Embrionarias Humanas/citología , Fosfatasa Alcalina/metabolismo , Blastocisto/citología , Línea Celular , Supervivencia Celular , Hibridación Genómica Comparativa , Genotipo , Prueba de Histocompatibilidad , Humanos , Masculino , Microscopía Fluorescente , Persona de Mediana EdadRESUMEN
The KCL039 human embryonic stem cell line was derived from a normal healthy blastocyst donated for research. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment and under current Good Manufacturing Practice (cGMP) standards. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Asunto(s)
Células Madre Embrionarias Humanas/citología , Diferenciación Celular , Línea Celular , Hibridación Genómica Comparativa , Fibroblastos/citología , Genotipo , Prueba de Histocompatibilidad , Células Madre Embrionarias Humanas/metabolismo , Humanos , Masculino , Microscopía Fluorescente , Persona de Mediana Edad , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
The KCL040 human embryonic stem cell line was derived from a normal healthy blastocyst donated for research. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment and under current Good Manufacturing Practice (cGMP) standards. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Asunto(s)
Células Madre Embrionarias Humanas/citología , Diferenciación Celular , Línea Celular , Hibridación Genómica Comparativa , Fibroblastos/citología , Genotipo , Prueba de Histocompatibilidad , Células Madre Embrionarias Humanas/metabolismo , Humanos , Masculino , Microscopía Fluorescente , Persona de Mediana Edad , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
The KCL021 human embryonic stem cell line was derived from an embryo donated for research that carried a ΔF508 mutation affecting the CFTR gene encoding the cystic fibrosis transmembrane conductance regulator. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Asunto(s)
Regulador de Conductancia de Transmembrana de Fibrosis Quística/genética , Células Madre Embrionarias Humanas/citología , Fosfatasa Alcalina/metabolismo , Diferenciación Celular , Línea Celular , Hibridación Genómica Comparativa , Embrión de Mamíferos/citología , Genotipo , Prueba de Histocompatibilidad , Células Madre Embrionarias Humanas/metabolismo , Humanos , Masculino , Microscopía Fluorescente , Persona de Mediana Edad , Mutación , LinajeRESUMEN
The KCL029 human embryonic stem cell line was derived from an embryo donated for research that carried a c.814T>C mutation in the WAS gene, which is linked to the Wiskott-Aldrich syndrome, a rare, inherited, X-linked, recessive disease characterized by immune dysregulation and microthrombocytopenia. The line is also carrier for a mutation p.N1152H in the gene encoding the cystic fibrosis transmembrane conductance regulator CFTR. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Asunto(s)
Células Madre Embrionarias Humanas/citología , Proteína del Síndrome de Wiskott-Aldrich/genética , Fosfatasa Alcalina/metabolismo , Diferenciación Celular , Línea Celular , Hibridación Genómica Comparativa , Regulador de Conductancia de Transmembrana de Fibrosis Quística/genética , Embrión de Mamíferos/citología , Genotipo , Prueba de Histocompatibilidad , Células Madre Embrionarias Humanas/metabolismo , Humanos , Masculino , Microscopía Fluorescente , Persona de Mediana Edad , Linaje , Polimorfismo de Nucleótido Simple , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
The KCL031 human embryonic stem cell line was derived from a normal healthy blastocyst donated for research. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment and under current Good Manufacturing Practice (cGMP) standards. Pluripotent state and differentiation potential were confirmed by in vitro and in vivo assays.
Asunto(s)
Células Madre Embrionarias Humanas/citología , Fosfatasa Alcalina/metabolismo , Blastocisto/citología , Diferenciación Celular , Línea Celular , Hibridación Genómica Comparativa , Genotipo , Prueba de Histocompatibilidad , Células Madre Embrionarias Humanas/metabolismo , Humanos , Cariotipo , Masculino , Microscopía Fluorescente , Persona de Mediana EdadRESUMEN
The KCL035 human embryonic stem cell line was derived from an embryo donated for research that carried a mutation in the HBB gene, which is linked to the ß-thalassemia syndrome. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Asunto(s)
Hemoglobinas Anormales/genética , Células Madre Embrionarias Humanas/citología , Fosfatasa Alcalina/metabolismo , Diferenciación Celular , Línea Celular , Hibridación Genómica Comparativa , Femenino , Fertilización In Vitro , Genotipo , Células Madre Embrionarias Humanas/metabolismo , Humanos , Cariotipo , Repeticiones de Microsatélite/genética , Microscopía Fluorescente , Linaje , Factores de Transcripción/metabolismoRESUMEN
The KCL024 human embryonic stem cell line was derived from an embryo donated for research that carried an autosomal dominant mutation in the NF1 gene encoding neurofibromin (c.3739-3742 ∆TTTG). Mutations in this gene have been linked to neurofibromatosis type 1, juvenile myelomonocytic leukemia and Watson syndrome. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Asunto(s)
Células Madre Embrionarias Humanas/citología , Neurofibromina 1/genética , Diferenciación Celular , Células Cultivadas , Embrión de Mamíferos/citología , Fertilización In Vitro , Prueba de Histocompatibilidad , Células Madre Embrionarias Humanas/metabolismo , Humanos , Microscopía Fluorescente , Neurofibromatosis 1/genética , Neurofibromatosis 1/patología , Linaje , Polimorfismo de Nucleótido Simple , Factores de Transcripción/metabolismoRESUMEN
The KCL026 human embryonic stem cell line was derived from an embryo donated for research that carried a mutation in the SMN1 gene encoding survival of motor neuron 1, telomeric (exons 7 and 8 deletion). Mutations in this gene are associated with spinal muscular atrophy. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Asunto(s)
Células Madre Embrionarias Humanas/citología , Proteína 1 para la Supervivencia de la Neurona Motora/genética , Diferenciación Celular , Células Cultivadas , Embrión de Mamíferos/citología , Exones , Fertilización In Vitro , Eliminación de Gen , Células Madre Embrionarias Humanas/metabolismo , Humanos , Masculino , Microscopía Fluorescente , Linaje , Factores de Transcripción/metabolismoRESUMEN
The KCL025 human embryonic stem cell line was derived from an embryo donated for research that carried an autosomal dominant mutation in the NF1 gene encoding neurofibromin (c.3739-3742 ΔTTTG). Mutations in this gene have been linked to neurofibromatosis type 1, juvenile myelomonocytic leukemia and Watson syndrome. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Asunto(s)
Células Madre Embrionarias Humanas/citología , Neurofibromina 1/genética , Diferenciación Celular , Células Cultivadas , Embrión de Mamíferos/citología , Fertilización In Vitro , Prueba de Histocompatibilidad , Células Madre Embrionarias Humanas/metabolismo , Humanos , Cariotipo , Masculino , Microscopía Fluorescente , Linaje , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
The KCL017 human embryonic stem cell line was derived from an embryo donated for research that carried an autosomal dominant mutation affecting splicing site of the VHL gene encoding von Hippel-Lindau tumor suppressor E3 ubiquitin protein ligase (676+3A>T). The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Asunto(s)
Células Madre Embrionarias Humanas/citología , Ubiquitina-Proteína Ligasas/genética , Diferenciación Celular , Células Cultivadas , Hibridación Genómica Comparativa , Embrión de Mamíferos/citología , Fertilización In Vitro , Genotipo , Células Madre Embrionarias Humanas/metabolismo , Humanos , Cariotipo , Masculino , Microscopía Fluorescente , Linaje , Polimorfismo Genético , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
The KCL013 human embryonic stem cell line was derived from an embryo donated for research that carried an autosomal dominant mutation affecting one allele of the HTT gene encoding huntingtin (42 trinucleotide repeats; 17 for the normal allele). The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Asunto(s)
Células Madre Embrionarias Humanas/citología , Proteína Huntingtina/genética , Alelos , Diferenciación Celular , Células Cultivadas , Reprogramación Celular , Hibridación Genómica Comparativa , Embrión de Mamíferos/citología , Fertilización In Vitro , Genotipo , Células Madre Embrionarias Humanas/metabolismo , Humanos , Cariotipo , Masculino , Microscopía Fluorescente , Linaje , Polimorfismo Genético , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Repeticiones de Trinucleótidos/genéticaRESUMEN
The KCL033 human embryonic stem cell line was derived from a normal healthy blastocyst donated for research. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment and under current Good Manufacturing Practice (cGMP) standards. Pluripotent state and differentiation potential were confirmed by in vitro assays. The line was also validated for sterility and specific and non-specific human pathogens.
Asunto(s)
Blastocisto/citología , Células Madre Embrionarias Humanas/citología , Diferenciación Celular , Células Cultivadas , Reprogramación Celular , Hibridación Genómica Comparativa , Femenino , Fertilización In Vitro , Genotipo , Prueba de Histocompatibilidad , Células Madre Embrionarias Humanas/metabolismo , Humanos , Cariotipo , Microscopía Fluorescente , Análisis de Secuencia por Matrices de Oligonucleótidos , Polimorfismo de Nucleótido Simple , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
The KCL034 human embryonic stem cell line was derived from a normal healthy blastocyst donated for research. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment and under current Good Manufacturing Practice (cGMP) standards. Pluripotent state and differentiation potential were confirmed by in vitro assays. The line was also validated for sterility, specific and non-specific human pathogens.
Asunto(s)
Células Madre Embrionarias Humanas/citología , Diferenciación Celular , Línea Celular , Hibridación Genómica Comparativa , Embrión de Mamíferos/citología , Genotipo , Prueba de Histocompatibilidad , Células Madre Embrionarias Humanas/metabolismo , Humanos , Masculino , Microscopía Fluorescente , Persona de Mediana Edad , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
The KCL012 human embryonic stem cell line was derived from an embryo donated for research that carried an autosomal dominant mutation affecting one allele of the HTT gene encoding huntingtin (46 trinucleotide repeats; 17 for the normal allele). The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment. Pluripotent state and differentiation potential were confirmed by in vitro and in vivo assays.