Abstract
Derivation of human embryonic stem cell (hESC) lines from chromosomally or genetically abnormal embryos obtained following preimplantation genetic diagnosis (PGD) is of immense interest to study various kinds of genetic disorders. In this study, we have established a new hESC line Relicell®hES4, isolated from an aneuploid embryo. Derivation of this cell line was achieved by isolation of the inner cell mass (ICM) by mechanical method. Karyotype analysis showed that the hESC line is euploid having 46 chromosomes, contrary to our expectations. The undifferentiated cells exhibited long-term proliferation capacity and expressed markers typical for hESC, such as OCT4, NANOG, and SSEA4. A comparative microarray study was carried out to analyze the transcription profile of Relicell®hES4 along with three other normal hESC line generated earlier in our lab. Relicell®hES4 manifested pluripotent differentiation potential both in vivo and in vitro. The cells were also induced to form neurons, cardiomyocytes, and pancreatic β islets. The generation of a normal hESC line from an abnormal embryo points to the fact that even such embryos can be considered for deriving new hESC lines instead of discarding them. The data represented here are the first detailed report on characterization and differentiation of an Indian hESC line generated from a PGD analyzed embryo.
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Acknowledgments
The authors acknowledge Reliance Life Sciences Pvt Ltd (www.rellife.com), for providing the infrastructure, administrative and financial support to work on this project. The authors thank Dr. Harinarayan Rao for his help in conducting the teratoma experiments and the team members of Molecular Medicine group for the karyotype analysis.
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Editor: T. Okamoto
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Antibodies details used for immunofluorescence staining (DOCX 12 kb)
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Mandal, A., Mathew, S., Saha, D. et al. Establishment, characterization, and differentiation of a karyotypically normal human embryonic stem cell line from a trisomy-affected embryo. In Vitro Cell.Dev.Biol.-Animal 49, 15–26 (2013). https://doi.org/10.1007/s11626-012-9567-z
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DOI: https://doi.org/10.1007/s11626-012-9567-z