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fetal cell isolation

Steen Kølvraa, Ripudaman Singh, Elizabeth A Normand, Sadeem Qdaisat, Ignatia B Van denVeyver, Laird Jackson, Lotte Hatt, Palle Schelde, Niels Uldbjerg, Else Marie Vestergaard, Li Zhao, Rui Chen, Chad A Shaw, Amy M Breman, Arthur L Beaudet
OBJECTIVE: Non-invasive prenatal testing (NIPT) based on fetal cells in maternal blood has the advantage over NIPT based on circulating cell-free fetal DNA in that there is no contamination with maternal DNA. This will most likely result in better detection of chromosomal aberrations including subchromosomal defects. The objective of this study was to test whether fetal cells enriched from maternal blood can be used for cell-based NIPT. METHODS: We present a method for enriching fetal cells from maternal blood, subsequent amplification of the fetal genome and detection of chromosomal and subchromosomal variations in the genome...
October 19, 2016: Prenatal Diagnosis
Vipulkumar Patel, Peter Celec, Magdalena Grunt, Heidi Schwarzenbach, Ingo Jenneckens, Timo Hillebrand
Circulating cell-free DNA (ccfDNA) is a promising diagnostic tool and its size fractionation is of interest. However, kits for isolation of ccfDNA available on the market are designed for small volumes hence processing large sample volumes is laborious. We have tested a new method that enables enrichment of ccfDNA from large volumes of plasma and subsequently allows size-fractionation of isolated ccfDNA into two fractions with individually established cut-off levels of ccfDNA length. This method allows isolation of low-abundant DNA as well as separation of long and short DNA molecules...
2016: Advances in Experimental Medicine and Biology
Emilio A Herrera, Francisca Cifuentes-Zúñiga, Esteban Figueroa, Cristian Villanueva, Cherie Hernández, René Alegría, Viviana Arroyo, Estefania Peñaloza, Marcelo Farías, Ricardo Uauy, Paola Casanello, Bernardo J Krause
In humans, intrauterine growth restriction (IUGR) is associated with vascular dysfunction, oxidative stress and signs of endothelial programming in umbilical vessels. We aimed to determine the effects of maternal antioxidant treatment with N-acetyl cysteine (NAC) on fetal endothelial function and eNOS programming in IUGR guinea pigs. IUGR was induced by implanting ameroid constrictors on uterine arteries of pregnant guinea pigs at mid gestation, receiving half of the sows NAC in the drinking water (from day 34 until term)...
October 14, 2016: Journal of Physiology
Azadeh Esmaeli, Mojgan Moshrefi, Ali Shamsara, Seyed Hasan Eftekhar-Vaghefi, Seyed Noureddin Nematollahi-Mahani
BACKGROUND: Fetal bovine serum (FBS) is widely used in cell culture laboratories, risk of zoonotic infections and allergic side effects create obstacles for its use in clinical trials. Therefore, an alternative supplement with proper inherent growth-promoting activities is demanded. OBJECTIVE: To find FBS substitute, we tested human umbilical cord blood serum (hUCS) for proliferation of human umbilical cord matrix derived mesenchymal stem cells (hUC-MSCs) and human bone marrow-derived mesenchymal cells (hBM-MSCs)...
September 2016: International Journal of Reproductive Biomedicine (Yazd, Iran)
Erica Sarchielli, Paolo Comeglio, Roberta Squecco, Lara Ballerini, Tommaso Mello, Giulia Guarnieri, Eglantina Idrizaj, Benedetta Mazzanti, Linda Vignozzi, Pasquale Gallina, Mario Maggi, Gabriella B Vannelli, Annamaria Morelli
CONTEXT: Previous studies have suggested that inflammatory pathways may impair central regulatory networks involving gonadotropin-releasing hormone (GnRH) neuron activity. Studies in humans are strongly hampered by the lack of human GnRH neuron cell lines. OBJECTIVE: To establish an in vitro model of GnRH neurons of human origin and analyze the effects of proinflammatory cytokines on their biological properties. Design, samples and treatments: The primary human fetal hypothalamic (hfHypo) cell cultures were isolated from brain of three 12 week-old fetuses...
October 13, 2016: Journal of Clinical Endocrinology and Metabolism
Florentine F Thurik, Marije Lamain-de Ruiter, Ahmad Javadi, Anneke Kwee, Heleen Woortmeijer, Godelieve C M L Page-Christiaens, Arie Franx, C Ellen van der Schoot, Wendy Koster
OBJECTIVE: To study associations of first trimester cell-free fetal DNA levels (in this paper referred to as cell-free placental DNA (cfpDNA) levels) and preeclampsia (PE), pregnancy-induced hypertension (PIH), gestational diabetes (GDM) and spontaneous preterm birth (sPB). METHOD: A nested case-control study was conducted in first trimester samples (gestational age (GA) 8(+0) -13(+6) weeks). A total of 226 cases and 301 controls were included. CfpDNA levels were quantified in male-bearing pregnancies using real-time DYS14-PCRs on DNA isolated from maternal serum...
October 12, 2016: Prenatal Diagnosis
Cassy M Spiller, Guillaume Burnet, Josephine Bowles
The fetal gonad contains a great variety of differentiating cell populations, of which germ cells make up a small percentage. In order to study germ cell-specific gene and protein expression, as well as determine direct effects of signaling molecules, it is necessary to prepare enriched populations of germ cells and maintain them in culture for several hours to multiple days. The protocols in this chapter are designed to provide a guide for the isolation or enrichment of mouse primordial germ cells (from 9...
2017: Methods in Molecular Biology
Jezamine Lim, Zainul Rashid Mohamad Razi, Jiaxian Law, Azmawati Mohammed Nawi, Ruszymah Binti Haji Idrus, Min Hwei Ng
BACKGROUND AIMS: Human Wharton's jelly-derived mesenchymal stromal cells (hWJMSCs) are possibly the most suitable allogeneic cell source for stromal cell therapy and tissue engineering applications because of their hypo-immunogenic and non-tumorigenic properties, easy availability and minimal ethical concerns. Furthermore, hWJMSCs possess unique properties of both adult mesenchymal stromal cells and embryonic stromal cells. The human umbilical cord (UC) is approximately 50-60 cm long and the existing studies in the literature have not provided information on which segment of the UC was studied...
October 7, 2016: Cytotherapy
Jairo A Diaz, Mauricio F Murillo, Jhonan A Mendoza, Ana M Barreto, Lina S Poveda, Lina K Sanchez, Laura C Poveda, Katherine T Mora
Emergent biological responses develop via unknown processes dependent on physical collision. In hypoxia, when the tissue architecture collapses but the geometric core is stable, actin cytoskeleton filament components emerge, revealing a hidden internal order that identifies how each molecule is reassembled into the original mold, using one common connection, i.e., a fractal self-similarity that guides the system from the beginning in reverse metamorphosis, with spontaneous self-assembly of past forms that mimics an embryoid phenotype...
2016: American Journal of Stem Cells
Khaled A Shibany, Sabine Tötemeyer, Stefanie L Pratt, Stuart W Paine
Despite reports of the successful isolation of primary equine hepatocytes, there are no published data regarding the successful cryopreservation of these isolated cells. In this study, a detailed description of the procedures for isolation, cryopreservation, and recovery of equine hepatocytes are presented. Furthermore, the intrinsic clearance (Clint) and production of metabolites for three drugs were compared between freshly isolated and recovered cryopreserved hepatocytes. Primary equine hepatocytes were isolated using a two-step collagenase perfusion method, with an average cell yield of 2...
October 2016: Pharmacology Research & Perspectives
Tereza C Cardoso, Lucas H Okamura, Jamila C Baptistella, Roberto Gameiro, Helena L Ferreira, Márcia Marinho, Eduardo F Flores
The possibility of isolating bovine mesenchymal multipotent stromal cells (MSCs) from fetal adnexa is an interesting prospect due to the potential use of these cells in biotechnological applications. However, little is known about the properties of these progenitor cells in bovine species. Wharton's jelly (WJ) MSC cells were obtained from the umbilical cord of bovine fetuses at three different stages of pregnancy and divided into groups 1, 2 and 3 according to gestational trimester. Cell morphology, from the three stages of pregnancy, typically appeared fibroblast-like spindle-shaped, presenting the same viability and number...
September 27, 2016: Cell and Tissue Research
Tally Lerman-Sagie, Zvi Leibovitz
Abnormal fetal corticogenesis results in malformations of cortical development (MCD). Abnormal cell proliferation leads to microcephaly or megalencephaly, incomplete neuronal migration results in heterotopia and lissencephaly, neuronal overmigration manifests as cobblestone malformations, and anomalous postmigrational cortical organization is responsible for polymicrogyria and focal cortical dysplasias. MCD comprises various congenital brain disorders, caused by different genetic, infectious, or vascular etiologies and is associated with significant neurological morbidity...
September 2016: Canadian Journal of Neurological Sciences. le Journal Canadien des Sciences Neurologiques
Briana Foley, Daniel Doheny, Michael Black, Salil N Pendse, Barbara Wetmore, Rebecca Clewell, Melvin E Andersen, Chad Deisenroth
The developmental origins of obesity hypothesis posits a multifaceted contribution of factors to the fetal origins of obesity and metabolic disease. Adipocyte hyperplasia in gestation and early childhood may result in predisposition for obesity later in life. Rodent in vitro and in vivo studies indicate that some chemicals may directly affect adipose progenitor cell differentiation, but the human relevance of these findings is unclear. The nuclear receptor peroxisome proliferator-activated receptor gamma (PPARG) is the master regulator of adipogenesis...
September 23, 2016: Toxicological Sciences: An Official Journal of the Society of Toxicology
Shiwen Liu, Chao Li, Yiyan Wang, Tingting Hong, Tiantian Song, Linxi Li, Leping Ye, Qingquan Lian, Ren-Shan Ge
The objective of the present study is to determine whether in utero exposure to methoxychlor (MXC) affects rat fetal Leydig cell number, cell size, or functions. Pregnant Sprague Dawley dams were gavaged with corn oil (control, 0mg/kg/day MXC) or MXC at doses of 10, 50, or 100mg/kg/day from gestational day (GD) 12 to 21. The results show that MXC increased fetal Leydig cell numbers dose-dependently from 95±8×10(3) cells/testis (control, mean±SEM) to 101±6, 148±22, and 168±21×10(3) cells/testis, at the doses of 10, 50, and 100mg/kg, respectively...
September 20, 2016: Toxicology
Michael J Burke
Seneca Valley Virus isolate 001 (SVV-001) is an oncolytic RNA virus of the Picornaviridae family. It is also the first picornavirus discovered of the novel genus Senecavirus. SVV-001 replicates through an RNA intermediate, bypassing a DNA phase, and is unable to integrate into the host genome. SVV-001 was originally discovered as a contaminant in the cell culture of fetal retinoblasts and has since been identified as a potent oncolytic virus against tumors of neuroendocrine origin. SVV-001 has a number of features that make it an attractive oncolytic virus, namely, its ability to target and penetrate solid tumors via intravenous administration, inability for insertional mutagenesis, and being a self-replicating RNA virus with selective tropism for cancer cells...
2016: Oncolytic Virotherapy
D-C Liu, X-N Yang, C-Z Huang, Y-G Sun, X-M Dai
OBJECTIVE: To observe whether adipose-derived stem cells (ADSCS), co-cultured with osteoblasts, can differentiate into osteoblasts and, if so, to study the best-induced conditions, with an ultimate goal of repairing bone defects. MATERIALS AND METHODS: Adipose-derived stem cells and osteoblasts were isolated from New Zealand white rabbits, and co-cultured in media with either 5% or 10% fetal bovine serum, for up to 4 weeks. The morphology of collected cells was examined under a microscope, and histological staining with alkaline phosphatase and alizarin red was carried out after induction for 1, 2, 3 and 4 weeks...
September 2016: European Review for Medical and Pharmacological Sciences
Matthew S Alexander, Anete Rozkalne, Alessandro Colletta, Janelle M Spinazzola, Samuel Johnson, Fedik Rahimov, Hui Meng, Michael W Lawlor, Elicia Estrella, Louis M Kunkel, Emanuela Gussoni
Cell-surface markers for prospective isolation of stem cells from human skeletal muscle have been difficult to identify. Such markers would be powerful tools for studying satellite cell function during homeostasis and in pathogenesis of diseases such as muscular dystrophies. In this study, we show that the tetraspanin KAI/CD82 is an excellent marker for prospectively isolating stem cells from human fetal and adult skeletal muscle. Human CD82(+) muscle cells robustly engraft into a mouse model of muscular dystrophy...
September 7, 2016: Cell Stem Cell
Jessica Aijia Liu, Martin Cheung
The neural crest (NC) is a remarkable transient structure generated during early vertebrate development. The neural crest progenitors have extensive migratory capacity and multipotency, harboring stem cell-like characteristics such as self-renewal. They can differentiate into a variety of cell types from craniofacial skeletal tissues to the trunk peripheral nervous system (PNS). Multiple regulators such as signaling factors, transcription factors, and migration machinery components are expressed at different stages of NC development...
September 14, 2016: Developmental Biology
Takako Tabata, Matthew Petitt, June Fang-Hoover, Martin Zydek, Lenore Pereira
Human cytomegalovirus (HCMV) is the leading viral cause of birth defects, including microcephaly, neurological deficits, hearing impairment, and vision loss. We previously reported that epithelial cells in amniotic membranes of placentas from newborns with intrauterine growth restriction and underlying congenital HCMV infection contain viral proteins in cytoplasmic vesicles. Herein, we immunostained amniotic membranes from 51 placentas from symptomatic and asymptomatic congenital infection with HCMV DNA in amniotic fluid and/or newborn saliva, intrauterine growth restriction, preterm deliveries, and controls...
September 13, 2016: American Journal of Pathology
Peck Yean Tan, Cheng Wei Chang, Kaibo Duan, Michael Poidinger, Kai Lyn Ng, Yap Seng Chong, Peter D Gluckman, Walter Stünkel
Wharton's jelly-derived Mesenchymal Stem Cells (MSCs) isolated from newborns with intrauterine fetal growth restriction were previously shown to exert anabolic features including insulin hypersensitivity. Here, we extend these observations and demonstrate that MSCs from small for gestational age (SGA) individuals have decreased mitochondrial oxygen consumption rates. Comparing normally grown and SGA MSCs using next generation sequencing studies, we measured global transcriptomic and epigenetic profiles and identified E2F1 as an over-expressed transcription factor regulating oxidative metabolism in the SGA group...
2016: PloS One
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