Gender Determination and Detection of Aneuploidy in Single Cells Using DNA Array-Based Comparative Genomic Hybridization

Hu, Dong; Guan, Xin Yuan; Hussey, N

doi:10.1007/978-1-59745-298-4_12

Cited by 8 publications

(2 citation statements)

References 20 publications

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“…Compared with other ARTs, the protocol required for PGD includes not only superovulation and embryo culture in vitro but also the more invasive biopsy procedure removing one or two blastomeres from the embryo. At present, research on PGD mostly focuses on making technical advancements to improve the accuracy and sensitivity of existing techniques and to develop new applications (Wilton, 2002; Wells and Levy, 2003; Hu et al, 2004, 2007). In contrast, relatively little evaluation of the influence of PGD on offspring development has been done, although the potential effects of in vitro fertilization (IVF) or intracytoplasmic sperm injection of eggs (ICSI) have attracted more attention.…”

Section: Introductionmentioning

confidence: 99%

Abnormal Development at Early Postimplantation Stage in Mouse Embryos After Preimplantation Genetic Diagnosis

Yang

Zeng

et al. 2012

The Anatomical Record

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Preimplantation genetic diagnosis (PGD) is an established procedure for the genetic analysis of embryos. To assess the effect of the procedure on early embryonic development, we generated a murine experimental system, including mice implanted with biopsied in vitro cultured embryos, control mice implanted with in vitro cultured embryos without biopsy, and mice with naturally conceived embryos. Embryos at the 7.5-dpc stage were isolated from all three groups and the embryo implantation rate, the survival rate of implanted embryos, and the developmental stage of surviving embryos were carefully assessed and compared among all three groups. We found the implantation rate was similar between biopsied and control group embryos (67.92% vs. 66.67%). However, the survival rate of implanted embryos in the biopsied group (49.31%) was significantly lower than that of the control (60.91%) and normal groups (96.24%) at 7.5 dpc. In addition, the survival rate of control group embryos was significant lower than that of normal group embryos. Classification of the precise developmental stages of randomly selected live implanted embryos at 7.5 dpc revealed no differences among the three groups. Our results indicate that blastomere biopsy does not adversely affect embryo implantation. The PGD procedure, in particular blastomere biopsy, increases the rate of embryo death at 4.5-7.5 dpc, but does not affect the development of surviving 7.5 dpc embryos.

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Section: Introductionmentioning

confidence: 99%

Abnormal Development at Early Postimplantation Stage in Mouse Embryos After Preimplantation Genetic Diagnosis

Yang

Zeng

et al. 2012

The Anatomical Record

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“…Further downstream, this enables robust molecular characterization and detection of chromosomal abnormalities [21,22]. Although different types of chromosomal abnormalities have been successfully identified by array-based comparative genomic hybridization (aCGH) following WGA of a small number of cells, the current aCGH protocol is a costly and time-consuming process that does not fit easily into all clinical schedules, particularly if specimens are required to be shipped to a reference laboratory [23,24,25,26,27]. Looking for an alternative approach that is clinically practical and has the potential to detect chromosomal abnormalities and single-gene disorders, the present study focuses on evaluating the fidelity of WGA using DNA obtained from a small number of fetal cells for a rapid and cost-effective prenatal diagnosis by multiplex quantitative fluorescent polymerase chain reaction (QF-PCR) [28].…”

Section: Introductionmentioning

confidence: 99%

Rapid Aneuploidy Detection of Chromosomes 13, 18, 21, X and Y Using Quantitative Fluorescent Polymerase Chain Reaction with Few Microdissected Fetal Cells

et al. 2015

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Objectives: Analysis of DNA from small numbers of cells, such as fetal cells in maternal blood, is a major limiting factor for their use in clinical applications. Traditional methods of single-cells whole genome amplification (SCs-WGA) and accurate analysis have been challenging to date. Our purpose was to assess the feasibility of using a few fetal cells to determine fetal sex and major chromosomal abnormalities by quantitative fluorescent polymerase chain reaction (QF-PCR). Methods: Cultured cells from 26 amniotic fluid samples were used for standard DNA extraction and recovery of 5 fetal cells by laser-capture microdissection. SCs-WGA was performed using the DNA from the microdissected cells. PCR amplification of short tandem repeats specific for chromosomes 13, 18, 21, X and Y was performed on extracted and amplified DNA. Allele dosage and sexing were quantitatively analyzed following separation by capillary electrophoresis. Results: Microsatellite QF-PCR analysis showed high concordance in chromosomal copy number between extracted and amplified DNA when 5 or more cells were used. Results were in concordance with that of conventional cytogenetic analysis. Conclusion: Satisfactory genomic coverage can be obtained from SCs-WGA. Clinically, SCs-WGA coupled with QF-PCR can provide a reliable, accurate, rapid and cost-effective method for detection of major fetal chromosome abnormalities.

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Preimplantation genetic diagnosis (PGD) influences adrenal development and response to cold stress in resulting mice

Zeng

et al. 2013

Cell Tissue Res

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Preimplantation genetic diagnosis (PGD) has gained widespread application in clinical medicine and hence the health of PGD offspring needs to be systematically assessed. Given the critical role of the stress response in growth and health, assessments of the development and function of the stress system might help to clarify the health outcomes of PGD. In this study, we constructed a PGD-conceived mouse model and used naturally conceived mice as controls; we used this model to evaluate the potential effect of PGD procedures on the stress system of the offspring. Serum and tissues of stress organs, namely the hypothalamus, locus coeruleus and adrenal gland, were collected from 5-week-old mice in the basal state or after cold stress. The serum levels of stress-related hormones and the structural and functional indices of the stress organs were then examined. In the basal state, ultrastructural abnormalities and low expression of genes involved in steroid hormone synthesis were found in the adrenals of the PGD mice, which had low corticosterone and high epinephrine levels compared with those of control mice. After acute cold stress, the PGD mice continued to show structural and glucocorticoid secretion abnormalities resulting in a late response to the environmental change. Thus, our study indicates that PGD manipulations affect adrenal development, result in structural and functional abnormalities of the adrenals in the offspring and influence their reactivity and adaptability to cold stress.

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Gender Determination and Detection of Aneuploidy in Single Cells Using DNA Array-Based Comparative Genomic Hybridization

Cited by 8 publications

References 20 publications

Abnormal Development at Early Postimplantation Stage in Mouse Embryos After Preimplantation Genetic Diagnosis

Abnormal Development at Early Postimplantation Stage in Mouse Embryos After Preimplantation Genetic Diagnosis

Rapid Aneuploidy Detection of Chromosomes 13, 18, 21, X and Y Using Quantitative Fluorescent Polymerase Chain Reaction with Few Microdissected Fetal Cells

Preimplantation genetic diagnosis (PGD) influences adrenal development and response to cold stress in resulting mice

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