TY - JOUR
T1 - Adaptation of Human Testicular Niche Cells for Pluripotent Stem Cell and Testis Development Research
AU - Pryzhkova, Marina V.
AU - Jordan, Philip W.
N1 - Funding Information:
We thank Immanuel Rasool from WRTC for coordinating the acquisition of deidentified human testis samples used for this study, Stephen Wellard for technical assistance, Ran Brosh for providing the EF1α promoter sequence, and Barry Zirkin and JinYong Chung for discussion. This work was supported by a KY Cha Award In Stem Cell Technology from the American Society for Reproductive Medicine and a research grant from National Institute of General Medical Sciences (R01GM11755), both awarded to P.W.J.
Funding Information:
We thank Immanuel Rasool from WRTC for coordinating the acquisition of deidentified human testis samples used for this study, Stephen Wellard for technical assistance, Ran Brosh for providing the EF1? promoter sequence, and Barry Zirkin and JinYong Chung for discussion. This work was supported by a KY Cha Award In Stem Cell Technology from the American Society for Reproductive Medicine and a research grant from National Institute of General Medical Sciences (R01GM11755), both awarded to P.W.J.
Publisher Copyright:
© 2020, The Korean Tissue Engineering and Regenerative Medicine Society.
PY - 2020/4/1
Y1 - 2020/4/1
N2 - Background:: Human testicular cells are greatly valuable to the research community as tools for studying testicular physiology and the effects of environmental pollutants. Because adult testicular cells have a limited self-organization capacity and life span, we investigated whether human pluripotent stem cells (hPSCs) can be used together with testicular cells to move a step closer toward making an optimal model of the human testis. Methods:: We used in vitro culture of donor testicular cells under serum-containing and chemically defined conditions. CRISPR-Cas9 technology was applied to introduce fluorescent transgenes (mCherry2 and EGFP) into hPSCs and testicular cells. hPSC-derived spheroids were co-cultured with human testicular cells in mini-spin bioreactors. Results:: Traditional cell culture conditions used for maintenance of testicular somatic cells generally contain serum and pose limitations on evaluating the role of active molecules on cell functions. We established that chemically defined culture conditions can be used to maintain testicular cells without the loss of proliferative activity. These cultures demonstrate marker expression which is characteristic of common testicular cell types: Sertoli, Leydig, endothelial, myoid cells, and macrophages. In order to model testicular physiology, it is important to be able to perform live cell microscopy. Thus, we generated fluorescent protein-expressing human testicular cells and hPSCs and demonstrated that these cell types can be successfully co-cultured for prolonged periods of time in a three-dimensional microenvironment. Conclusion:: Our research extends the possible applications of human testis-derived somatic cells and shows that they can be used together with hPSCs for further studies of human male reproductive biology.
AB - Background:: Human testicular cells are greatly valuable to the research community as tools for studying testicular physiology and the effects of environmental pollutants. Because adult testicular cells have a limited self-organization capacity and life span, we investigated whether human pluripotent stem cells (hPSCs) can be used together with testicular cells to move a step closer toward making an optimal model of the human testis. Methods:: We used in vitro culture of donor testicular cells under serum-containing and chemically defined conditions. CRISPR-Cas9 technology was applied to introduce fluorescent transgenes (mCherry2 and EGFP) into hPSCs and testicular cells. hPSC-derived spheroids were co-cultured with human testicular cells in mini-spin bioreactors. Results:: Traditional cell culture conditions used for maintenance of testicular somatic cells generally contain serum and pose limitations on evaluating the role of active molecules on cell functions. We established that chemically defined culture conditions can be used to maintain testicular cells without the loss of proliferative activity. These cultures demonstrate marker expression which is characteristic of common testicular cell types: Sertoli, Leydig, endothelial, myoid cells, and macrophages. In order to model testicular physiology, it is important to be able to perform live cell microscopy. Thus, we generated fluorescent protein-expressing human testicular cells and hPSCs and demonstrated that these cell types can be successfully co-cultured for prolonged periods of time in a three-dimensional microenvironment. Conclusion:: Our research extends the possible applications of human testis-derived somatic cells and shows that they can be used together with hPSCs for further studies of human male reproductive biology.
KW - Genetic modification
KW - Human pluripotent stem cells
KW - Human testicular cells
KW - Mini-spin bioreactor
KW - Organoids
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U2 - 10.1007/s13770-020-00240-0
DO - 10.1007/s13770-020-00240-0
M3 - Article
C2 - 32114677
AN - SCOPUS:85080993242
SN - 1738-2696
VL - 17
SP - 223
EP - 235
JO - Tissue Engineering and Regenerative Medicine
JF - Tissue Engineering and Regenerative Medicine
IS - 2
ER -