Chromosome Preparation From Cultured Cells

Howe, Bradley; Umrigar, Ayesha; Tsien, Fern

doi:10.3791/50203-v

Cited by 9 publications

(10 citation statements)

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“…Karyotyping was performed using peripheral blood samples. Phytohemagglutinin culture was established and harvested after 72 h. Twenty metaphases were then analyzed following Giemsa‐Trypsin banding 22 …”

Section: Methodsmentioning

confidence: 99%

“…Phytohemagglutinin culture was established and harvested after 72 h. Twenty metaphases were then analyzed following Giemsa-Trypsin banding. 22 For exome sequencing, genomic DNA of an affected family member (IV-2) was selected. The DNA was prepared using a SureSelect Target Enrichment Kit (Agilent, Santa Clara, CA, USA) in accordance with the manufacturer's instructions.…”

Section: Analysis Of Semen and Reproductive Hormones (Luteinizing Hor...mentioning

confidence: 99%

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A variant in sperm‐specific glycolytic enzyme enolase 4 (ENO4) causes human male infertility

Nawaz,

Hussain,

Bilal

et al. 2023

The Journal of Gene Medicine

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BackgroundAlthough defects in sperm morphology and physiology lead to male infertility, in many instances, the exact disruption of molecular pathways in a given patient is often unknown. The glycolytic pathway is an essential process to supply energy in sperm cell motility. Enolase 4 (ENO4) is crucial for the glycolytic process, which provides the energy for sperm cells in motility. ENO4 is located in the sperm principal piece and is essential for the motility and organization of the sperm flagellum. In the present study, we characterized a family with asthenozoospermia and abnormal sperm morphology as a result of a variant in the enolase 4 (ENO4) gene.MethodsComputer‐assisted semen analysis, papanicolaou smear staining and scanning electron microscopy were used to examine sperm motility and morphology for semen analysis in patients. For genetic analysis, whole‐exome sequencing followed by Sanger sequencing was performed.ResultsTwo brothers in a consanguineous family were being clinically investigated for sperm motility and morphology issues. Genetic analysis by whole‐exome sequencing revealed a homozygous variant [c.293A>G, p.(Lys98Arg)] in the ENO4 gene that segregated with infertility in the family, shared by affected but not controls.ConclusionsIn view of the association of asthenozoospermia and abnormal sperm morphology in Eno4 knockout mice, we consider this to be the first report describing the involvement of ENO4 gene in human male infertility. We also explore the possible involvement of another variant in explaining other phenotypic features in this family.

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Section: Methodsmentioning

confidence: 99%

Section: Analysis Of Semen and Reproductive Hormones (Luteinizing Hor...mentioning

confidence: 99%

A variant in sperm‐specific glycolytic enzyme enolase 4 (ENO4) causes human male infertility

Nawaz,

Hussain,

Bilal

et al. 2023

The Journal of Gene Medicine

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“…The peripheral blood samples of proband and her parents were obtained for cytogenetic analysis of karyotype and for dna extraction [using the Magna Pure 96 System (roche diagnostics, ltd.) according to the manufacturer's recommendations] for molecular cytogenetic/genetic analyses. The cytogenetic analysis of proband's karyotype was performed using the routine G-banding procedure by Giemsa-romanowski staining (7). The whole-genome screening of submicroscopic copy-number variations (cnVs) and copy-number neutral losses of heterozygosity (cnnloH) was performed using the oligonucleotide-based microarray platform SurePrint G3 iSca v2 cGH+SnP 4X180K Microarray following the manufacturer's recommendations (agilent Technologies, inc.).…”

Section: Methodsmentioning

confidence: 99%

A unique case of Bloom syndrome with a combination of genetic hits: A lesson from trio‑based exome sequencing: A case report

et al. 2023

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Pathogenic variants affecting the BLM gene are responsible for the manifestation of extremely rare cancer-predisposing Bloom syndrome. The present study reports on a case of an infant with a congenital hypotrophy, short stature and abnormal facial appearance. initially she was examined using a routine molecular diagnostic algorithm, including the cytogenetic analysis of her karyotype, microarray analysis and methylation-specific MLPA, however, she remained undiagnosed on a molecular level. Therefore, she and her parents were enrolled in the project of trio-based exome sequencing (eS) using Human core exome kit. She was revealed as a carrier of an extremely rare combination of causative sequence variants altering the BLM gene (nM_000057.4), c.1642c>T and c.2207_2212delinsTaGaTTc in the compound heterozygosity, resulting in a diagnosis of Bloom syndrome. Simultaneously, a mosaic loss of heterozygosity of chromosome 11p was detected and then confirmed as a borderline imprinting center 1 hypermethylation on chromosome 11p15. The diagnosis of Bloom syndrome and mosaic copy-number neutral loss of heterozygosity of chromosome 11p increases a lifetime risk to develop any types of malignancy. This case demonstrates the trio-based eS as a complex approach for the molecular diagnostics of rare pediatric diseases.

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“…Chromosome analysis was done using G-banding analysis (400 bands) (10). As much as 3 ml of peripheral venous blood was inserted into a venoject containing sodium heparin.…”

Section: Cell Culture Methodsmentioning

confidence: 99%

Chromosome Aberration on Growth and Developmental Disorder

Setijowati

Suprapti

Sugeng

et al. 2022

JKB

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Growth and development in humans begin at conception, which shows progress and interdependence. Normal children show the characteristic of growth and development. In children with growth and development disorder, chromosomal abnormality may be found. This study aimed to determine chromosomal abnormality in patients with growth and development disorder. The samples were taken from karyotype results of patients in the Medical Genetics Laboratory, Faculty of Medicine, Universitas Wijaya Kusuma Surabaya, from 2010-2020. The inclusion criteria were infants to adolescents aged 24 years and unmarried who experienced impaired growth and development, including sexual development. Chromosomal abnormalities were obtained from cytogenetic analysis using the G-banding method. From 75 samples with growth and development disorders, there were abnormalities in the number of autosomal chromosomes and sex chromosomes found in patients with Down syndrome, Turner syndrome, Klinefelter syndrome, and Edwards syndrome. Chromosomal structural abnormalities found were deletion, translocation, inversion, duplication, marker chromosome, and heteromorphism. This study showed the importance of karyotyping in children and adolescents with growth and developmental disorders.

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Chromosome Preparation From Cultured Cells

Cited by 9 publications

References 0 publications

A variant in sperm‐specific glycolytic enzyme enolase 4 (ENO4) causes human male infertility

A variant in sperm‐specific glycolytic enzyme enolase 4 (ENO4) causes human male infertility

A unique case of Bloom syndrome with a combination of genetic hits: A lesson from trio‑based exome sequencing: A case report

Chromosome Aberration on Growth and Developmental Disorder

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