# Establishment of human mini-Testis for reproductive toxicity testing

> **NIH NIH R43** · REPROTOX BIOTECH, LLC · 2020 · $225,000

## Abstract

Summary
 Occupational exposures during manufacture and application of these chemicals represent the highest
level of exposure that could cause potential adverse health effects. There remains a considerable knowledge gap
regarding the relationship between workplace exposure of industrial chemicals and possible adverse health
effects. Current test methods set by regulatory agencies predominately rely on animal testing. The cost and
time with the animal testing for risk assessment seriously limit efforts for extensive chemical screening. These
data gaps necessitate the implementation of a new vision for toxicity testing as reflected in the National
Research Council’s report “Toxicity Testing in the 21st Century”. Under the support of NIOSH R21 OH010473
grant, we established a rodent testicular cell co-culture model and found this in vitro co-culture model could
identify the reproductive toxicants. While results from these in vivo or in vitro rodent models are informative,
these models could not recapitulate the whole process of human spermatogenesis as rodent spermatogenesis
proceeds in a distinctly different fashion compared to humans. Thus, developing in vitro human models for
reproductive toxicity testing is paramount for advancing the field and for understanding the cellular and
molecular mechanisms that may underlie these disruptions in human spermatogenesis. In this proposal, we
will develop a human testicular cell co-culture model, and demonstrate the reliability and sensitivity of this
model in high-throughput and multi-parametric high-content analytical screening for workplace chemicals on
the reproductive and developmental systems. The specific aims are (1) to establish testicular cell co-culture
model from human spermatogonial stem cells (hSSCs), Sertoli cell and Leydig cells; (2) to develop integrated
assays based on the adverse outcome pathways; (3) to apply the multi-parametric high-throughput and high-
content analysis (HT/HCA) to examine and compare reproductive toxic and non-toxic compounds to validate
the in vitro testicular cell co-culture model. Through this hypothesis-driven research, we will test the
hypothesis that an in vitro 3D human testicular cell co-culture model creates a proper microenvironment
that mimics a human testis to promote the growth of germ cells in vitro and generate reproducible results for
predicting reproductive toxicity. The research proposed will result in publications and generate toxicity data of
the chemicals in the workplace, and provide data for risk assessment. Therefore, this proposal meets the
mission of Cancer, Reproductive, and Cardiovascular Research Program (CRC). The identification of
reproductive toxicants will reduce hazardous exposures and lower the risk of reproductive
malfunction. Therefore, our proposed project is within the NIOSH CRC’s Research to Practice (r2p)
strategy to promote the transfer and translation of research findings into prevention practices.

## Key facts

- **NIH application ID:** 10010729
- **Project number:** 1R43ES031890-01
- **Recipient organization:** REPROTOX BIOTECH, LLC
- **Principal Investigator:** Lei Yin
- **Activity code:** R43 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $225,000
- **Award type:** 1
- **Project period:** 2020-08-01 → 2023-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10010729

## Citation

> US National Institutes of Health, RePORTER application 10010729, Establishment of human mini-Testis for reproductive toxicity testing (1R43ES031890-01). Retrieved via AI Analytics 2026-05-28 from https://api.ai-analytics.org/grant/nih/10010729. Licensed CC0.

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