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• W2022119918 abstract "OBJECTIVE: Since many frequent, fatal, and incurable diseases do not have an appropriate disease model and with attainable embryos being scarce, with IRB approval we attempted to generate novel disease-specific human embryonic stem (hES) cells with new protocols for more efficient derivation, maintenance, and differentiation.DESIGN: Experimental.MATERIALS AND METHODS: Zona-free human blastocysts (n=14) previously assessed by preimplantation genetic diagnosis (PGD) for genetic conditions were transferred onto feeder cells and cultured in DMEM-based media. Pieces of the colonies (n=4) were frozen by liquid nitrogen vitrification with cryoprotectants propylene glycol, DMSO, and acetamide and subsequently thawed. Differentiation of hES cells was achieved by colony overgrowth or embryoid body formation. Embryonic and control cells were subjected to marker gene and protein analysis for pluripotency and differentiation by reverse transcription PCR and immunofluorescence, respectively.RESULTS: Colonies (n=10; 71.4% of transferred blastocysts) could be established and maintained which showed the typical morphological features of hES cells such as compact colony formation. Colonies were derived from affected embryos (one each of cystic fibrosis, trisomy X, 18, 21, 22, and Tay-Sachs disease), from embryos tested inconclusively (one of cystic fibrosis and three of Tay-Sachs disease), and from three normal control embryos. Marker gene and protein expression as well as growth pattern analysis suggest that the colony cells retain their undifferentiated state in culture as well as after vitrification and thawing and that they can be differentiated into a variety of cell types, including the tissues most affected by the conditions.CONCLUSIONS: These newly established protocols for the derivation, maintenance, and differentiation of novel disease-specific hES cell lines should enable the efficient generation of new disease models. This will provide new tools to study diseases as well as to develop new therapeutic approaches. OBJECTIVE: Since many frequent, fatal, and incurable diseases do not have an appropriate disease model and with attainable embryos being scarce, with IRB approval we attempted to generate novel disease-specific human embryonic stem (hES) cells with new protocols for more efficient derivation, maintenance, and differentiation. DESIGN: Experimental. MATERIALS AND METHODS: Zona-free human blastocysts (n=14) previously assessed by preimplantation genetic diagnosis (PGD) for genetic conditions were transferred onto feeder cells and cultured in DMEM-based media. Pieces of the colonies (n=4) were frozen by liquid nitrogen vitrification with cryoprotectants propylene glycol, DMSO, and acetamide and subsequently thawed. Differentiation of hES cells was achieved by colony overgrowth or embryoid body formation. Embryonic and control cells were subjected to marker gene and protein analysis for pluripotency and differentiation by reverse transcription PCR and immunofluorescence, respectively. RESULTS: Colonies (n=10; 71.4% of transferred blastocysts) could be established and maintained which showed the typical morphological features of hES cells such as compact colony formation. Colonies were derived from affected embryos (one each of cystic fibrosis, trisomy X, 18, 21, 22, and Tay-Sachs disease), from embryos tested inconclusively (one of cystic fibrosis and three of Tay-Sachs disease), and from three normal control embryos. Marker gene and protein expression as well as growth pattern analysis suggest that the colony cells retain their undifferentiated state in culture as well as after vitrification and thawing and that they can be differentiated into a variety of cell types, including the tissues most affected by the conditions. CONCLUSIONS: These newly established protocols for the derivation, maintenance, and differentiation of novel disease-specific hES cell lines should enable the efficient generation of new disease models. This will provide new tools to study diseases as well as to develop new therapeutic approaches." @default.
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• W2022119918 date "2009-09-01" @default.
• W2022119918 modified "2023-09-26" @default.
• W2022119918 title "Novel highly efficient generation of disease-specific human embryonic stem cells from genetically abnormal embryos" @default.
• W2022119918 doi "https://doi.org/10.1016/j.fertnstert.2009.07.332" @default.
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