title: Up-regulation of heme oxygenase-1 expression modulates reactive oxygen species level during the cryopreservation of human seminiferous tubules.
abstract: OBJECTIVE
  To study the effect of freezing techniques and to optimize a method for trace amounts of testicular spermatozoa from biopsed seminiferous tubules. The level of reactive oxygen species (ROS) and the gene expression of heme oxygenase-1 was evaluated.
  
  
  METHODS
  Prospective experimental study.
  
  
  METHODS
  University-based laboratory.
  
  
  METHODS
  Eighteen adults with male fator infertility underwent testicular biopsy surgery.
  
  
  METHODS
  Seminiferous tubular fragments from each man were evenly allocated to three groups: fresh control, slow freezing, and vitrifiaction groups. The morphology and ROS levels before and after freezing were evaluated for seminiferous tubular fragments. The expression of heme oxygenase-1 (HO-1) at both the transcriptional and protein levels was determined.
  
  
  METHODS
  The morphology was analyzed by light microscopy. The ROS levels were measured with ELISA. The proliferation and differentiation were evaluated by immunohistochemistry, and the expression of HO-1 was evaluated using a real-time polymerase chain reaction (PCR) and Western blotting.
  
  
  RESULTS
  Decreased ROS levels and increased HO-1 expression at the transcriptional and protein levels were observed after thawing the human seminiferous tubules. The ROS level was negatively correlated with HO-1 expression. Slow freezing was more effective than vitrification in terms of HO-1 up-regulation and ROS alteration.
  
  
  CONCLUSIONS
  Based on our study, the slow freezing technique was more effective compared with the vitrification method.
authors.affiliation: Department of Urology, Peking University Third Hospital, Beijing, People's Republic of China; Key Laboratory of Assisted Reproduction, Ministry of Education, Beijing, People's Republic of China.
authors.forename: Wenhao
authors.initials: W
authors.lastname: Tang
authors.affiliation: Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, People's Republic of China; Key Laboratory of Assisted Reproduction, Ministry of Education, Beijing, People's Republic of China.
authors.forename: Jie
authors.initials: J
authors.lastname: Yan
authors.affiliation: Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, People's Republic of China; Key Laboratory of Assisted Reproduction, Ministry of Education, Beijing, People's Republic of China.
authors.forename: Tianren
authors.initials: T
authors.lastname: Wang
authors.affiliation: Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, People's Republic of China; Key Laboratory of Assisted Reproduction, Ministry of Education, Beijing, People's Republic of China.
authors.forename: Xi
authors.initials: X
authors.lastname: Xia
authors.affiliation: Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, People's Republic of China; Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproduction, Beijing, People's Republic of China.
authors.forename: Xinjie
authors.initials: X
authors.lastname: Zhuang
authors.affiliation: Department of Urology, Peking University Third Hospital, Beijing, People's Republic of China; Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproduction, Beijing, People's Republic of China.
authors.forename: Kai
authors.initials: K
authors.lastname: Hong
authors.affiliation: Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, People's Republic of China; Key Laboratory of Assisted Reproduction, Ministry of Education, Beijing, People's Republic of China.
authors.forename: Rong
authors.initials: R
authors.lastname: Li
authors.affiliation: Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, People's Republic of China; Key Laboratory of Assisted Reproduction, Ministry of Education, Beijing, People's Republic of China.
authors.forename: Ping
authors.initials: P
authors.lastname: Liu
authors.affiliation: Department of Urology, Peking University Third Hospital, Beijing, People's Republic of China; Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproduction, Beijing, People's Republic of China.
authors.forename: Hui
authors.initials: H
authors.lastname: Jiang
authors.affiliation: Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, People's Republic of China; Key Laboratory of Assisted Reproduction, Ministry of Education, Beijing, People's Republic of China. Electronic address: jie.qiao@263.net.
authors.forename: Jie
authors.initials: J
authors.lastname: Qiao
mesh_terms: D001706:Biopsy; D002454:Cell Differentiation; D049109:Cell Proliferation; D048430:Cell Shape; D015925:Cryopreservation / Q000379:methods*; D015972:Gene Expression Regulation, Neoplastic; D051547:Heme Oxygenase-1 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008297:Male; D018384:Oxidative Stress; D011446:Prospective Studies; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012671:Seminiferous Tubules / Q000201:enzymology* / Q000473:pathology; D013094:Spermatozoa / Q000201:enzymology / Q000473:pathology; D013997:Time Factors; D015854:Up-Regulation; D058989:Vitrification
chemical_list: D017382:Reactive Oxygen Species; C496541:HMOX1 protein, human; D051547:Heme Oxygenase-1
keywords: Oxidative stress; cryopreservation; testicular spermatozoa; trace amounts