In vitro spermatogenesis model for assessing male reproductive toxicity

Details

• Personal Author:
  Liang S ; Wei H ; Yin L ; Yu X
• Description:
  Reproductive toxicity testing in animals represents one of the largest uses of animals. It has been an extraordinarily challenging area to implement in vitro alternatives due to the complexity of reproductive systems. We previously developed a three-dimensional testicular cells co-culture from rat testis and validated this in vitro model can discriminate developmentally toxic phthalate esters (PEs) from non-toxic PEs. However, this model still employs a large number of animals for isolation of primary testicular cells. The goal of this study is to establish a testicular cells co-culture system (3D-TCCS) from the testicular cell lines including spermatogonial stem cells C18-4, Sertoli cell TM4 and Leydig cell TM3. We examined both the morphology and cellular biomarkers of this novel in vitro 3D-TCCS model. In an effort to establish a toxicity-based high throughput model, we compared the dose-dependent effects on cell viability of known male reproductive toxic PEs (DPP, DBP, BBP and DEHP) and non-toxic PEs (DEP, DMP and DOTP). Cell viability was measured by Neutral Red assays at 24 and 48 h. DBP, DEHP, BBP and DPP treatments were found to result in dose-dependent decreases in cell viability. Those observations are consistent with the fact that these four PEs are classified as developmentally toxic. The advantage of this 3D-TCCS allows further to clarify the target cells by examining the cellular responses in an individual cell culture model. We found that DBP and BBP targeted on both spermatogonia C18-4 cells and TM3 cells, while DPP targeted all three types of cells. DEHP was found to target both the TM3 and TM4 cells while DEP, DMP and DOTP only induced decrease of cell viability at the highest concentration of 400 micoM in the TM4 cells. All these data demonstrated that this animal free in vitro model has the potential to offer predictive modeling for assessing male reproductive toxicity without sacrifice of animals. We are currently further developing an integrated pathway HCA and HTP screening assay in the 3D-TCCS model for male reproductive toxicity evaluation. [Description provided by NIOSH]
• Subjects:
  Men Occupational Health Phthalic Acids Safety Spermatogenesis Toxicology Urogenital System
• Keywords:
  Dose Response In Vitro Study Phthalates Reproductive Effects Testes
• ISSN:
  1096-6080
• Document Type:
  Text
• Funding:
  Grant
• Genre:
  Abstract or Summary
• Place as Subject:
  California ; Georgia ; OSHA Region 4 ; OSHA Region 9
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Volume:
  144
• Issue:
  1
• NIOSHTIC Number:
  nn:20046038
• Citation:
  Toxicologist 2015 Mar; 144(1):250
• Federal Fiscal Year:
  2015
• Performing Organization:
  University of Georgia, Athens
• Peer Reviewed:
  False
• Start Date:
  20130901
• Source Full Name:
  The Toxicologist. Society of Toxicology 54th Annual Meeting and ToxExpo, March 22-26, 2015, San Diego, California
• End Date:
  20160831
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:5e4c698b21962d62628cf00fd881c8eb7102d82579615246ad3cb37d58e512cad8f76ec5532a2f60ec5609917c986d23e13b658347afbe6f178220cd74d424f6
• Download URL:
  https://stacks.cdc.gov/view/cdc/201223/cdc_201223_DS1.pdf
• File Type:
  [PDF - 1.07 MB ]