title: Optimization of techniques for multiple platform testing in small, precious samples such as human chorionic villus sampling.
abstract: BACKGROUND
  Multiple testing to understand global changes in gene expression based on genetic and epigenetic modifications is evolving. Chorionic villi, obtained for prenatal testing, is limited, but can be used to understand ongoing human pregnancies. However, optimal storage, processing and utilization of CVS for multiple platform testing have not been established.
  
  
  RESULTS
  Leftover CVS samples were flash-frozen or preserved in RNAlater. Modifications to standard isolation kits were performed to isolate quality DNA and RNA from samples as small as 2-5 mg. RNAlater samples had significantly higher RNA yields and quality and were successfully used in microarray and RNA-sequencing (RNA-seq). RNA-seq libraries generated using 200 versus 800-ng RNA showed similar biological coefficients of variation. RNAlater samples had lower DNA yields and quality, which improved by heating the elution buffer to 70 °C. Purification of DNA was not necessary for bisulfite-conversion and genome-wide methylation profiling. CVS cells were propagated and continue to express genes found in freshly isolated chorionic villi.
  
  
  CONCLUSIONS
  CVS samples preserved in RNAlater are superior. Our optimized techniques provide specimens for genetic, epigenetic and gene expression studies from a single small sample which can be used to develop diagnostics and treatments using a systems biology approach in the prenatal period. © 2016 John Wiley & Sons, Ltd.
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Cedars-Sinai Medical Center, Los Angeles, CA, USA.|Department of Obstetrics and Gynecology, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.
authors.forename: Margareta D
authors.initials: MD
authors.lastname: Pisarska
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Cedars-Sinai Medical Center, Los Angeles, CA, USA.
authors.forename: Marzieh
authors.initials: M
authors.lastname: Akhlaghpour
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Cedars-Sinai Medical Center, Los Angeles, CA, USA.
authors.forename: Bora
authors.initials: B
authors.lastname: Lee
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Cedars-Sinai Medical Center, Los Angeles, CA, USA.
authors.forename: Gillian M
authors.initials: GM
authors.lastname: Barlow
authors.affiliation: Division of Endocrinology, Diabetes and Metabolism, Department of Medicine, Cedars-Sinai Medical Center, Los Angeles, CA, USA.
authors.forename: Ning
authors.initials: N
authors.lastname: Xu
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Cedars-Sinai Medical Center, Los Angeles, CA, USA.|Department of Obstetrics and Gynecology, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.
authors.forename: Erica T
authors.initials: ET
authors.lastname: Wang
authors.affiliation: Center for Public Health Genomics, University of Virginia, Charlottesville, VA, USA.
authors.forename: Aaron J
authors.initials: AJ
authors.lastname: Mackey
authors.affiliation: Center for Public Health Genomics, University of Virginia, Charlottesville, VA, USA.
authors.forename: Charles R
authors.initials: CR
authors.lastname: Farber
authors.affiliation: Center for Public Health Genomics, University of Virginia, Charlottesville, VA, USA.
authors.forename: Stephen S
authors.initials: SS
authors.lastname: Rich
authors.affiliation: Institute for Translational Genomics and Population Sciences, LABiomed/Harbor-UCLA Medical Center, Torrance, CA, USA.
authors.forename: Jerome I
authors.initials: JI
authors.lastname: Rotter
authors.affiliation: Institute for Translational Genomics and Population Sciences, LABiomed/Harbor-UCLA Medical Center, Torrance, CA, USA.
authors.forename: Yii-der I
authors.initials: YI
authors.lastname: Chen
authors.affiliation: Division of Endocrinology, Diabetes and Metabolism, Department of Medicine, Cedars-Sinai Medical Center, Los Angeles, CA, USA.
authors.forename: Mark O
authors.initials: MO
authors.lastname: Goodarzi
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT, USA.
authors.forename: Seth
authors.initials: S
authors.lastname: Guller
authors.affiliation: Department of Obstetrics and Gynecology, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.|Division of Maternal Fetal Medicine, Department of Obstetrics and Gynecology, Cedars-Sinai Medical Center, Los Angeles, CA, USA.
authors.forename: John
authors.initials: J
authors.lastname: Williams
mesh_terms: D018929:Cell Culture Techniques; D015193:Chorionic Villi Sampling; D019411:Clinical Laboratory Techniques; D004247:DNA / Q000302:isolation & purification; D005260:Female; D006801:Humans; D011247:Pregnancy; D012313:RNA / Q000302:isolation & purification
chemical_list: D012313:RNA; D004247:DNA