Engineering and characterization of simplified vaginal and seminal fluid simulants

Rachna Rastogi, Jonathan Su, Alamelu Mahalingam, Justin Clark, Samuel Sung, Thomas Hope, Patrick F. Kiser*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

25 Scopus citations


Background Reported vaginal and seminal fluid simulants have complex compositions with multiple preparatory steps that contribute to physical instability. We report the design and characterization of stable and simplified buffers that mimic the salient physical/chemical properties of the physiological fluids. Study design/methods Human cervicovaginal and seminal fluid samples were collected and buffering capacity was determined. The major buffering species were identified from published compositions of reproductive tract fluids. These values were used to compute the composition of vaginal and seminal fluid simulants. Ionic strength, buffering capacities, pH and osmolalities were then calculated or experimentally determined. Finally, cytotoxicity was evaluated in HEC-1-A cells and 3D reconstructed EpiVaginal™ tissue (VEC-100-FT) using naïve cells/tissue and nonoxynol-9 as controls. Results The use of calculated amounts of conjugate acid and base for buffer development resulted in compositions that did not require endpoint pH adjustment and could be formulated as stable 10 × concentrates. Furthermore, due to the absence of complex divalent salts, all our proposed simulants were stable at 4°C for 1 month whereas precipitation and pH and osmolality changes were noted in reported buffers. Experimental determination of buffering capacities yielded similar values for undiluted cervicovaginal fluid (β4.2-5.2= 35.6±12.3 mM, N= 7) and human seminal fluid (β7 -6= 37.5±5 mM, N= 3). All neat simulants showed significant cytotoxicity in HEC-1-A cells but were well tolerated by organotypic vaginal tissue. Conclusions We report revised and improved compositions of buffers mimicking salient properties of vaginal and seminal fluid necessary for in vitro product evaluation. Implications To support research in reproductive health and in particular drug delivery, we have designed and characterized stable new media to mimic these important fluids that can be used in a variety of in vitro studies.

Original languageEnglish (US)
Pages (from-to)337-346
Number of pages10
Issue number4
StatePublished - Apr 1 2016


  • Buffering capacity
  • Cytotoxicity
  • Human cervicovaginal fluid
  • Simulant design

ASJC Scopus subject areas

  • Obstetrics and Gynecology
  • Reproductive Medicine


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