scholarly journals Construction of amniotic cell line with chromosomal abnormalities and its application in the quality control of chromosome karyotype analysis

2019 ◽  
Vol 33 (1) ◽  
pp. 33-37
Author(s):  
Binghuan Weng ◽  
Jun Ying ◽  
Hanzhi Wang ◽  
Danhu Zhu ◽  
Lanjuan Li
Keyword(s):  
Quality Control ◽  
Cell Line ◽  
Chromosomal Abnormalities ◽  
Karyotype Analysis ◽  
Amniotic Cell ◽  
Chromosome Karyotype

scholarly journals Combined Application of Chromosome Karyotype and Microarray Analysis in Fetus With Increased Nuchal Translucency

2021 ◽  
Author(s):  
Wang Chaohong ◽  
Tang Junxiang ◽  
Sun Yuxiu ◽  
Pang Yu ◽  
Tong Keting ◽  
...  
Keyword(s):  
Prenatal Diagnosis ◽  
Pregnant Women ◽  
Microarray Analysis ◽  
Chromosomal Abnormalities ◽  
Karyotype Analysis ◽  
Nuchal Translucency ◽  
Chromosome Abnormalities ◽  
Detection Rates ◽  
Combined Application ◽  
Chromosome Karyotype

Abstract Objective：To examine the risk of chromosomal abnormalities when the thickness of the nuchal translucency( NT ) is 2.5-2.9mm, to evaluate the cutoff value of NT for prenatal diagnosis, to explore the value of combined application of chromosome karyotype and microarray analysis, and to explore the relationship between NT ≥ 5.0mm and fetal prognosis. Methods：A total of 366 pregnant women who underwent prenatal diagnosis in Anhui Province Maternity and Child Health Hospital from January 2018 to August 2020 were collected, of which 241 cases had NT ≥ 2.5mm, 125 cases were elderly (35-38 years old) with NT < 2.5mm. We made grouping statistics on chromosome abnormalities,and compared the detection of chromosome abnormalities by karyotype and microarray analysis. At the same time, we followed up the fetuses with NT ≥ 5.0mm and analyzed their prognosis. Results: (1)Among the 366 cases with NT thickening,the detection rates of chromosome abnormalities by karyotype analysis and microarray analysis(CMA) were 13.39% (49/366) and 13.93% (51/366), respectively, and there was no significant difference (P>0.05), including 25 cases of trisomy 21, 5 cases of trisomy 18, 5 cases of Turner synthesis and 16 cases of other chromosome abnormalities. (2)We compared the effect of different NT value on the detection rate of pathogenic chromosomes, and found that the difference between NT ≥2.5mm and NT < 2.5mm was statistically significant(P<0.05). The detection rates of pathogenic chromosomal abnormalities in NT < 2.5mm group,2.5-2.9mm group, 3.0-3.4mm group,3.5-4.4mm group,4.5-5.4mm group and NT ≥ 5.5mm group were 0.8%(1/ 125), 11.63%(10/ 86), 17.81%(13/ 73), 20%(10/ 49), 47.62%(10/ 21) and 63.64%(7/ 11) respectively. (3)Our study found that different prenatal diagnostic indicators for abnormal chromosome detection rate difference was statistically significant(P<0.05). The detection rates of NT thickening alone and NT thickening combined with other abnormalities were 13.17%(22/ 167) and 35.14%(26/ 74) respectively(P<0.05). (4)Among 18 pregnant women with NT ≥ 5.0 mm, 9 fetuses were chromosomal abnormalities, and 9 fetuses survived healthily. Conclusions：When the NT value is 2.5-2.9mm, the incidence of fetal chromosome abnormality is significantly higher than that in the normal group. It is suggested that invasive prenatal diagnosis should be performed for pregnant women with NT ≥ 2.5mm. Chromosome karyotype analysis and CMA can complement each other, which is conducive to prenatal genetic counseling. The fetuses with NT thickening usually have good pregnancy outcomes when excluding fetal chromosome and prenatal ultrasound does not indicate any abnormalities.

scholarly journals Tribological, biocompatibility, and antibiofilm properties of tungsten–germanium coating using magnetron sputtering

2021 ◽  
Vol 32 (1) ◽  
Author(s):  
Mustafa Şükrü Kurt ◽  
Mehmet Enes Arslan ◽  
Ayşenur Yazici ◽  
İlkan Mudu ◽  
Elif Arslan
Keyword(s):  
Cell Culture ◽  
Magnetron Sputtering ◽  
Cell Line ◽  
Borosilicate Glass ◽  
Chromosomal Abnormalities ◽  
Dermal Fibroblast ◽  
Fibroblast Cell ◽  
Antibiofilm Activity ◽  
Borosilicate Glasses

AbstractIn this study, borosilicate glass and 316 L stainless steel were coated with germanium (Ge) and tungsten (W) metals using the Magnetron Sputtering System. Surface structural, mechanical, and tribological properties of uncoated and coated samples were examined using SEM, X-ray diffraction (XRD), energy-dispersive spectroscopy, and tribometer. The XRD results showed that WGe2 chemical compound observed in (110) crystalline phase and exhibited a dense structure. According to the tribological analyses, the adhesion strength of the coated deposition on 316 L was obtained 32.8 N, and the mean coefficient of friction was around 0.3. Biocompatibility studies of coated metallic biomaterials were analyzed on fibroblast cell culture (Primary Dermal Fibroblast; Normal, Human, Adult (HDFa)) in vitro. Hoescht 33258 fluorescent staining was performed to investigate the cellular density and chromosomal abnormalities of the HDFa cell line on the borosilicate glasses coated with germanium–tungsten (W–Ge). Cell viabilities of HDFa cell line on each surface (W–Ge coated borosilicate glass, uncoated borosilicate glass, and cell culture plate surface) were analyzed by using (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) cytotoxicity assay. The antibiofilm activity of W–Ge coated borosilicate glass showed a significant reduction effect on Staphylococcus aureus (ATCC 25923) and Pseudomonas aeruginosa (ATCC 27853) adherence compared to control groups. In the light of findings, tungsten and germanium, which are some of the most common industrial materials, were investigated as biocompatible and antimicrobial surface coatings and recommended as bio-implant materials for the first time.

The unfolded protein response in a dolichyl phosphate mannose-deficient Chinese hamster ovary cell line points out the key role of a demannosylation step in the quality-control mechanism of N-glycoproteins

2002 ◽  
Vol 362 (2) ◽  
pp. 491-498 ◽  
Author(s):  
François FOULQUIER ◽  
Anne HARDUIN-LEPERS ◽  
Sandrine DUVET ◽  
Ingrid MARCHAL ◽  
Anne Marie MIR ◽  
...  
Keyword(s):  
Quality Control ◽  
Control System ◽  
Unfolded Protein Response ◽  
Cell Line ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster ◽  
Quality Control System ◽  
Unfolded Protein ◽  
The Unfolded Protein Response ◽  
Protein Response

The CHO (Chinese hamster ovary) glycosylation mutant cell line, B3F7, transfers the truncated glycan Glc3Man5GlcNAc2 on to nascent proteins. After deglucosylation, the resulting Man5GlcNAc2 glycan is subjected to two reciprocal enzymic processes: the action of an endoplasmic-reticulum (ER) kifunensine-sensitive α1,2-mannosidase activity to yield a Man4GlcNAc2 glycan, and the reglucosylation involved in the quality-control system which ensures that only correctly folded glycoproteins leave the ER. We show that the recombinant secreted alkaline phosphatase (SeAP) produced in stably transfected B3F7 cells, is co-immunoprecipitated with the GRP78 (glucose-regulated protein 78), a protein marker of the unfolded protein response (UPR). The level of GRP78 transcription has been evaluated by reverse transcription-PCR (RT-PCR) and we demonstrate that B3F7 cells present a constitutively higher level of UPR in the absence of inductors, compared with Pro−5 cells. Interestingly, a decrease was observed in the UPR and an increase in SeAP secretion in the kifunensine-treated B3F7 cells. Altogether, these data highlight the relationships between the glycan structure, the quality control system and the UPR. Moreover, they support the idea that a specific demannosylation step is a key event of the glycoprotein quality control in B3F7 cells.

scholarly journals Clinical Utility and the Yield of Single Nucleotide Polymorphism Array in Prenatal Diagnosis of Fetal Central Nervous System Abnormalities

2021 ◽  
Vol 8 ◽  
Author(s):  
Meiying Cai ◽  
Hailong Huang ◽  
Liangpu Xu ◽  
Na Lin
Keyword(s):  
Central Nervous System ◽  
Single Nucleotide Polymorphism ◽  
Copy Number ◽  
Chromosomal Abnormalities ◽  
Karyotype Analysis ◽  
Snp Array ◽  
Copy Number Variations ◽  
Nucleotide Polymorphism ◽  
Single Nucleotide ◽  
Pathogenic Cnvs

Applying single nucleotide polymorphism (SNP) array to identify the etiology of fetal central nervous system (CNS) abnormality, and exploring its association with chromosomal abnormalities, copy number variations, and obstetrical outcome. 535 fetuses with CNS abnormalities were analyzed using karyotype analysis and SNP array. Among the 535 fetuses with CNS abnormalities, chromosomal abnormalities were detected in 36 (6.7%) of the fetuses, which were consistent with karyotype analysis. Further, additional 41 fetuses with abnormal copy number variations (CNVs) were detected using SNP array (the detection rate of additional abnormal CNVs was 7.7%). The rate of chromosomal abnormalities, but not that of pathogenic CNVs in CNS abnormalities with other ultrasound abnormalities was significantly higher than that in isolated CNS abnormalities. The rates of chromosomal abnormalities and pathogenic CNVs in fetuses with spine malformation (50%), encephalocele (50%), subependymal cyst (20%), and microcephaly (16.7%) were higher than those with other isolated CNS abnormalities. The pregnancies for 36 cases with chromosomal abnormalities, 18 cases with pathogenic CNVs, and three cases with VUS CNVs were terminated. SNP array should be used in the prenatal diagnosis of fetuses with CNS abnormalities, which can enable better prenatal assessment and genetic counseling, and affect obstetrical outcomes.

scholarly journals Keeping it clean: the cell culture quality control experience at the National Center for Advancing Translational Sciences

2019 ◽  
Author(s):  
Jacob S. Roth ◽  
Tobie D. Lee ◽  
Dorian M. Cheff ◽  
Maya L. Gosztyla ◽  
Rosita Asawa ◽  
...  
Keyword(s):  
Quality Control ◽  
Cell Line ◽  
Cell Lines ◽  
High Throughput Screening ◽  
Science Research ◽  
Rapid Identification ◽  
Contamination Rate ◽  
Mycoplasma Contamination ◽  
Systematic Testing ◽  
Short Tandem

AbstractQuality control monitoring of cell lines utilized in biomedical research is of critical importance, and is critical for reproducibility of data. Two key pitfalls in tissue culture are 1) cell line authenticity, and 2) mycoplasma contamination. As a collaborative research institute, the National Center for Advancing Translational Sciences receives cell lines from a range of commercial and academic sources, that are adapted for high-throughput screening. Here, we describe the implementation of a routine NCATS-wide mycoplasma testing and short-tandem repeat (STR) testing for cell lines. Initial testing identified a >10% mycoplasma contamination rate, but the implementation of clearly defined protocols that included immediate destruction of contaminated cell lines wherever possible has led to a much-reduced mycoplasma contamination rate, and data for >2,000 cell line samples tested over 3 years, and case studies are provided. STR testing of 170 cell lines with established STR profiles revealed only 5 mis-identified cell lines received from external labs. The data collected over the three years since implementation of this systematic testing demonstrates the importance of continual vigilance for rapid identification of ‘problem’ cell lines, for ensuring reproducible data in translational science research.

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