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Technique to ‘Map' Chromosomal Mosaicism at the Blastocyst Stage

Taylor, Tyl H., Griffin, Darren K., Katz, Seth L., Crain, Jack L., Johnson, Lauren, Gitlin, Susan (2016) Technique to ‘Map' Chromosomal Mosaicism at the Blastocyst Stage. Cytogenetic and Genome Research, 149 (4). pp. 262-266. ISSN 1424-8581. (doi:10.1159/000449051) (KAR id:58532)

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http://doi.org/10.1159/000449051

Abstract

The purpose of this study was to identify a technique that allows for comprehensive chromosome screening (CCS) of individual cells within human blastocysts along with the approximation of their location in the trophectoderm relative to the inner cell mass (ICM). This proof-of-concept study will allow for a greater understanding of chromosomal mosaicism at the blastocyst stage and the mechanisms by which mosaicism arises. One blastocyst was held by a holding pipette and the ICM was removed. While still being held, the blastocyst was further biopsied into quadrants. To separate the individual cells from the biopsied sections, the sections were placed in calcium/magnesium-free medium with serum for 20 min. A holding pipette was used to aspirate the sections until individual cells were isolated. Individual cells from each section were placed into PCR tubes and prepped for aCGH. A total of 18 cells were used for analysis, of which 15 (83.3%) amplified and provided a result and 3 (16.7%) did not. Fifteen cells were isolated from the trophectoderm; 13 (86.7%) provided an aCGH result, while 2 (13.3%) did not amplify. Twelve cells were euploid (46,XY), while 1 was complex abnormal (44,XY), presenting with monosomy 7, 10, 11, 13, and 19, and trisomy 14, 15, and 21. A total of 3 cells were isolated from the ICM; 2 were euploid (46,XY) and 1 did not amplify. Here, we expand on a previously published technique which disassociates biopsied sections of the blastocyst into individual cells. Since the blastocyst sections were biopsied in regard to the position of the ICM, it was possible to reconstruct a virtual image of the blastocyst while presenting each cell's individual CCS results.

Item Type: Article
DOI/Identification number: 10.1159/000449051
Subjects: Q Science
Divisions: Divisions > Division of Natural Sciences > School of Biosciences
Depositing User: Susan Davies
Date Deposited: 10 Nov 2016 16:43 UTC
Last Modified: 23 Jan 2020 04:12 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/58532 (The current URI for this page, for reference purposes)
Griffin, Darren K.: https://orcid.org/0000-0001-7595-3226
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