The Early History of Cell Culture
Howard Green in Therapy with Cultured Cells, 2019
Since the early 19th century, the scientific world has been acquainted with the study of microorganisms in culture. But in order to study animal cells, it was necessary to isolate them from the organism and keep them alive in vitro . The first well-documented account of the cultivation of animal tissue in vitro is that of Harrison, at Johns Hopkins University. He demonstrated that embryonic tissue of the frog would develop normally in an explant culture. A long period of time elapsed before attention was first directed to defining the nutritional needs of cultured cells, so as to eliminate the use of complex media containing embryo extracts. These investigators demonstrated that living tissue fragments could be dissociated with trypsin to produce cell suspensions that could be plated like bacteria, and that the cells could be subcultured at least twice. Cells obtained from mice, hamsters and other animals, when serially cultivated, become aneuploid and develop into established cell lines.
The Application of Genetic Tests in an Assisted Reproduction Unit: Sperm FISH
Nicolás Garrido, Rocio Rivera in A Practical Guide to Sperm Analysis, 2017
Intracytoplasmic sperm injection (ICSI) allows overcoming the natural barrier offered by the oocyte to sperm fertilization in couples with severe male factor infertility. Sperm fluorescence in situ hybridization (FISH) uses fluorescence DNA probes directed to specific DNA sequences in the interphase sperm nuclei. Visualizing the hybridization signals using fluorescence microscopy, it is possible to identify numerical chromosome abnormalities in the nucleus of ejaculated, epididymal, and testicular sperm. Sperm FISH analyses corroborate previous reports with altered meiosis in infertile men, as they have shown higher aneuploidy rates for chromosome 21 and sex chromosomes due to meiotic nondisjunction. Sperm heads have a tightly compacted nucleus due to the presence of disulfide bridges between protamines; this condensation of nuclear chromatin makes it inaccessible to DNA probes. Double-strand DNA denaturation of the sperm and FISH probes is carried out after incubation at high temperature. The hybridization signals are visualized using a fluorescent microscope equipped with specific filters for each fluorochrome.
Genetics
Karl H. Pang, Nadir I. Osman, James W.F. Catto, Christopher R. Chapple in Basic Urological Sciences, 2021
Genes are the basic physical unit of inheritance. Every person has two copies (alleles) of each gene — inherited from each parent. Aneuploidy is an abnormal number of chromosomes. Structural changes can cause problems with growth, development, and function. Clinical and phenotypic effects depend on their size, location, or gain/loss of genetic material. Structural changes include : Deletion: loss of genetic material, Duplication: genetic material is copied, Inversion: DNA breaks in two places and the resulting piece of DNA is reversed and reinserted, and Isochromosomes: two identical arms, instead of one short (p) and one long (q) arm. Only female ova contribute mitochondria to the developing embryo; therefore, only females can pass on mitochondrial mutations.
Aneuploidy Detection in Paraffin Embedded Tissue from Products of Conception by Mini-STR Genotyping
Published in Fetal and Pediatric Pathology, 2013
Larissa V. Furtado, Mohamed A. Jama, Christian N. Paxton, Andrew A. Wilson, Anna E. Gardiner, Elaine Lyon, Katherine B. Geiersbach
Autosomal trisomy is the most common genetic abnormality observed in pregnancy loss. We designed a panel of mini-short tandem repeats (mini-STRs) for aneuploidy detection in chromosomes 13, 16, 18 and 21 from fresh and formalin fixed, paraffin embedded (FFPE) samples from products of conception (POC). FFPE POCs with trisomy 13 (n = 6), trisomy 18 (n = 6), trisomy 21 (n = 12), 6 euploid for the chromosomes of interest and two trisomy 16 samples from fresh tissue were tested. Concordance between cytogenetics and genotyping was 100% for non-mosaic samples. Mini-STR genotyping is a viable method for targeted aneuploidy detection in low quality DNA samples.
The impact of inherited thrombophilia on first trimester combined aneuploidy screening test parameters
Published in The Journal of Maternal-Fetal & Neonatal Medicine, 2014
Mehmet Fatih Karsli, Eralp Baser, Kerem Doga Seckin, Mahmut İlkin Yeral, Cihan Togrul, Mustafa Ugur
Objective: To determine whether inherited thrombophilia affects components of first trimester combined aneuploidy screening test. Method: A case–control study was performed between January 1st and December 31st 2011, at a tertiary referral hospital. Singleton pregnancies with inherited thrombophilia that underwent first trimester (11–13+6 week) combined aneuploidy screening test were included in the study. Pregnancy associated plasma protein-A (PAPP-A), free beta-human chorionic gonadotropin (fbHCG) and fetal nuchal translucency (NT) were compared between the study group and controls. Results: Within the study period, 15 881 women with singleton pregnancies had a combined first trimester aneuploidy screening test at our institution. Among these, 207 women met the inclusion criteria. A control group that comprised 625 women with similar gestational age was generated, using a 1:3 ratio. PAPP-A levels were significantly higher, whereas fbHCG levels and fetal NT measurements were lower in women with inherited thrombophilia (p
A study of aneuploidy and DNA fragmentation in spermatozoa of three men with sex chromosome mosaicism including a 45,X cell line
Published in Human Fertility, 2015
Minh Huong Nguyen, Frederic Morel, Louis Bujan, Pascale May-Panloup, Marc De Braekeleer, Aurore Perrin
Meiotic segregation of mosaic males with a 45,X cell line has been little examined. In this study, we evaluated the risk of aneuploid gametes using fluorescence in situ hybridization (FISH) and DNA fragmentation in ejaculated spermatozoa of three men with sex chromosome mosaicism including a 45,X cell line. Triple- and dual-color FISH were performed. Sperm DNA fragmentation was detected using the TUNEL assay. A significantly increased frequency of XY disomic spermatozoa was observed for patients (P)1 and P2. A significant increase in diploidy and autosomal aneuploidy was found in P2 and P3, respectively. The rate of DNA fragmentation was not different from that observed in a control group. Data from the literature are scarce (only 3 cases reported), making comparison of the present data difficult, especially as the frequencies of the cell lines comprising the mosaicism differed between patients. Furthermore, the proportion of the different cell lines can differ from one tissue to another in the same patient. Whether the relative levels of the several cell lines present in the mosaicism can influence the rate of aneuploid spermatozoa remains unknown.