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High-efficiency derivation of human embryonic stem cell lines following pre-implantation genetic diagnosis.
- Source :
-
In vitro cellular & developmental biology. Animal [In Vitro Cell Dev Biol Anim] 2010 Apr; Vol. 46 (3-4), pp. 376-85. Date of Electronic Publication: 2010 Mar 09. - Publication Year :
- 2010
-
Abstract
- Pre-implantation genetic diagnosis allows the characterisation of embryos that carry a gene responsible for a severe monogenic disease and to transfer to the mother's uterus only the unaffected one(s). The genetically affected embryos can be used to establish human embryonic stem cell (hESC) lines. We are currently establishing a cell bank of ESC lines carrying specific disease-causing mutant genes. These cell lines are available to the scientific community. For this purpose, we have designed a technique that requires only minimal manipulation of the embryos. At the blastocyst stage, we just removed the zona pellucida before seeding the embryo as a whole on a layer of feeder cells. This approach gave a good success rate (>20%), whatever the quality of the embryos, and allowed us to derive 11 new hESC lines, representing seven different pathologies. Full phenotypic validation of the cell lines according to ISCI guidelines confirmed their pluripotent nature, as they were positive for hESC markers and able to differentiate in vitro in all three germ layers derivatives. Nine out of 11 stem cell lines had normal karyotypes. Our results indicate that inner cell mass isolation is not mandatory for hESC derivation and that minimal manipulation of embryos can lead to high success rate.
- Subjects :
- Animals
Antigens, Surface metabolism
Biomarkers metabolism
Cell Differentiation genetics
Cell Line
Female
Gene Expression Regulation, Developmental
Humans
Karyotyping
Male
Mice
Pedigree
Pluripotent Stem Cells cytology
Pluripotent Stem Cells metabolism
Reverse Transcriptase Polymerase Chain Reaction
Blastocyst cytology
Cell Culture Techniques methods
Embryonic Stem Cells cytology
Preimplantation Diagnosis methods
Subjects
Details
- Language :
- English
- ISSN :
- 1543-706X
- Volume :
- 46
- Issue :
- 3-4
- Database :
- MEDLINE
- Journal :
- In vitro cellular & developmental biology. Animal
- Publication Type :
- Academic Journal
- Accession number :
- 20217271
- Full Text :
- https://doi.org/10.1007/s11626-010-9300-8