Q3164493 (Q3164493): Difference between revisions
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(Created claim: summary (P836): Although it is relatively easy to generate induced pluripotential cells (iPSCs), its genetic aberrations and oncogenic potential limit its use in regenerative medicine. On the other hand, the generation of embryonic stem cells (ESCs) through nuclear transfer (NSTS) is very inefficient, requires a large number of good quality oocytes and, in humans, is associated with ethical and logistical limitations. Alternatively, several studies have shown t...) |
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Although it is relatively easy to generate induced pluripotential cells (iPSCs), its genetic aberrations and oncogenic potential limit its use in regenerative medicine. On the other hand, the generation of embryonic stem cells (ESCs) through nuclear transfer (NSTS) is very inefficient, requires a large number of good quality oocytes and, in humans, is associated with ethical and logistical limitations. Alternatively, several studies have shown that permeabilisation and incubation of somatic cells with oocyte extracts from both lower species (Xenopus, Drosophila) and mammals (mouse, pig) induce their reprogramming evidenced by the expression of pluripotence markers. However, this type of study also requires the use of a very high number of oocytes, being impracticable with samples of human origin. We propose the use of a microfluidic device to generate liposomes from oocyte extracts and direct the controlled fusion of individual liposomes to somatic cells. Our hypothesis is that the controlled fusion of a critical volume of oocyte extract into each somatic cell can induce its efficient reprogramming. The epigenetic and transcriptomic profile of oocyte-treated cells shall be compared to somatic cell profiles treated with liposomes without the corresponding extract, ESCs and iPSCs. We propose to optimise this system using mouse cells and then reprogram cells of human origin with homologous oocytes. The successful implementation of this platform will allow us to generate stem cells of human origin and the unprecedented opportunity to study the oocyte factors responsible for reprogramming. (English) | |||||||||||||||
Property / summary: Although it is relatively easy to generate induced pluripotential cells (iPSCs), its genetic aberrations and oncogenic potential limit its use in regenerative medicine. On the other hand, the generation of embryonic stem cells (ESCs) through nuclear transfer (NSTS) is very inefficient, requires a large number of good quality oocytes and, in humans, is associated with ethical and logistical limitations. Alternatively, several studies have shown that permeabilisation and incubation of somatic cells with oocyte extracts from both lower species (Xenopus, Drosophila) and mammals (mouse, pig) induce their reprogramming evidenced by the expression of pluripotence markers. However, this type of study also requires the use of a very high number of oocytes, being impracticable with samples of human origin. We propose the use of a microfluidic device to generate liposomes from oocyte extracts and direct the controlled fusion of individual liposomes to somatic cells. Our hypothesis is that the controlled fusion of a critical volume of oocyte extract into each somatic cell can induce its efficient reprogramming. The epigenetic and transcriptomic profile of oocyte-treated cells shall be compared to somatic cell profiles treated with liposomes without the corresponding extract, ESCs and iPSCs. We propose to optimise this system using mouse cells and then reprogram cells of human origin with homologous oocytes. The successful implementation of this platform will allow us to generate stem cells of human origin and the unprecedented opportunity to study the oocyte factors responsible for reprogramming. (English) / rank | |||||||||||||||
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Property / summary: Although it is relatively easy to generate induced pluripotential cells (iPSCs), its genetic aberrations and oncogenic potential limit its use in regenerative medicine. On the other hand, the generation of embryonic stem cells (ESCs) through nuclear transfer (NSTS) is very inefficient, requires a large number of good quality oocytes and, in humans, is associated with ethical and logistical limitations. Alternatively, several studies have shown that permeabilisation and incubation of somatic cells with oocyte extracts from both lower species (Xenopus, Drosophila) and mammals (mouse, pig) induce their reprogramming evidenced by the expression of pluripotence markers. However, this type of study also requires the use of a very high number of oocytes, being impracticable with samples of human origin. We propose the use of a microfluidic device to generate liposomes from oocyte extracts and direct the controlled fusion of individual liposomes to somatic cells. Our hypothesis is that the controlled fusion of a critical volume of oocyte extract into each somatic cell can induce its efficient reprogramming. The epigenetic and transcriptomic profile of oocyte-treated cells shall be compared to somatic cell profiles treated with liposomes without the corresponding extract, ESCs and iPSCs. We propose to optimise this system using mouse cells and then reprogram cells of human origin with homologous oocytes. The successful implementation of this platform will allow us to generate stem cells of human origin and the unprecedented opportunity to study the oocyte factors responsible for reprogramming. (English) / qualifier | |||||||||||||||
point in time: 12 October 2021
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Revision as of 16:34, 12 October 2021
Project Q3164493 in Spain
Language | Label | Description | Also known as |
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English | No label defined |
Project Q3164493 in Spain |
Statements
24,000.0 Euro
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48,000.0 Euro
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50.0 percent
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1 January 2015
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31 March 2019
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UNIVERSIDAD DE LERIDA
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25120
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Aunque es relativamente fácil generar células pluripotenciales inducidas (iPSCs), sus aberraciones genéticas y potencial oncogenicidad limitan su uso en medicina regenerativa. Por otra parte, la generación de células madre embrionarias (ESCs) a través de la transferencia nuclear (SCNT) es muy ineficiente, requiere un gran número de ovocitos de buena calidad y, en humanos, se asocia a limitaciones éticas y logísticas. Como alternativa, varios estudios han demostrado que la permeabilización e incubación de células somáticas con extractos de ovocito tanto de especies inferiores (Xenopus, Drosophila) como de mamíferos (ratón, cerdo) induce su reprogramación evidenciada por la expresión de marcadores de pluripotencia. No obstante, este tipo de estudios también precisan de la utilización de un número muy elevado de ovocitos, siendo impracticable con muestras de origen humano. Proponemos la utilización de un dispositivo microfluídico para generar liposomas a partir de extractos de ovocito y dirigir la fusión controlada de liposomas individuales a células somáticas. Nuestra hipótesis es que la fusión controlada de un volumen crítico de extracto de ovocito a cada célula somática puede inducir su eficiente reprogramación. El perfil epigenético y transcriptómico de las células tratadas con ovocitos se comparará a los perfiles de células somáticas tratadas con liposomas sin el correspondiente extracto, ESCs y iPSCs. Proponemos optimizar este sistema usando células de ratón para luego reprogramar células de origen humano con ovocitos homólogos. La implementación de esta plataforma con éxito nos permitirá generar células madre de origen humano y la oportunidad sin precedentes de estudiar los factores del ovocito responsables de la reprogramación. (Spanish)
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Although it is relatively easy to generate induced pluripotential cells (iPSCs), its genetic aberrations and oncogenic potential limit its use in regenerative medicine. On the other hand, the generation of embryonic stem cells (ESCs) through nuclear transfer (NSTS) is very inefficient, requires a large number of good quality oocytes and, in humans, is associated with ethical and logistical limitations. Alternatively, several studies have shown that permeabilisation and incubation of somatic cells with oocyte extracts from both lower species (Xenopus, Drosophila) and mammals (mouse, pig) induce their reprogramming evidenced by the expression of pluripotence markers. However, this type of study also requires the use of a very high number of oocytes, being impracticable with samples of human origin. We propose the use of a microfluidic device to generate liposomes from oocyte extracts and direct the controlled fusion of individual liposomes to somatic cells. Our hypothesis is that the controlled fusion of a critical volume of oocyte extract into each somatic cell can induce its efficient reprogramming. The epigenetic and transcriptomic profile of oocyte-treated cells shall be compared to somatic cell profiles treated with liposomes without the corresponding extract, ESCs and iPSCs. We propose to optimise this system using mouse cells and then reprogram cells of human origin with homologous oocytes. The successful implementation of this platform will allow us to generate stem cells of human origin and the unprecedented opportunity to study the oocyte factors responsible for reprogramming. (English)
12 October 2021
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Lleida
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Identifiers
PI14_02184
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