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Biofabricating Seminiferous Tubules for In Vitro Spermatogenesis

用于体外精子发生的生物制造曲细精管

基本信息

批准号：
10800970
负责人：
Yuguo Lei
金额：
$ 41.38万
依托单位：
PENNSYLVANIA STATE UNIVERSITY, THE
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-20 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10800970
关键词：
3-Dimensional Acids Affect Alginates Animal Testing Architecture Binding Biological Assay Blood-Testis Barrier Buffers Cell Culture System Cell Culture Techniques Cell Maintenance Cells Chemicals Crosslinker Cultured Cells Custom Diagnosis Diameter Diffusion Edetic Acid Endothelial Cells Epithelium Germ Cells Growth Homing Hormones Human Hydrogels In Vitro Infertility Ions Knock-out Knowledge Male Infertility Mammalian Cell Matrix Metalloproteinases Methods Modeling Mus Patients Peptides Phenotype Physiology Process Production Pump RGD (sequence)Reaction Research Risk Scientist Seminiferous tubule structure Shapes Side Spermatogenesis Spermatogonia Suspension Culture System Technology Testis Time Tissues Toxic effect Tube Tubular formation boys cell growth crosslink divinyl sulfone improved in vivo infertility treatment interstitial interstitial cell male mouse model new technology prepuberty prevent scaffold sertoli cell sperm cell stem cells

项目摘要

Project Summary Failed spermatogenesis is a major cause of male infertility. In vivo, spermatogenesis occurs in seminiferous tubules. The process is regulated by factors secreted by the interstitial tissue surrounding the tubules. An in vitro testis cell culture system mimics the seminiferous tubule architecture and models spermatogenesis would significantly advance fundamental studies and developing methods to prevent, diagnose, and treat infertility. In past decades, scientists have developed various testis cell culture systems, including 2D culture and 3D cell suspension culture with or without scaffold support for in vitro spermatogenesis (IVS). However, the cellular organization of seminiferous tubules has not been successfully established in vitro. Because the architecture is critical for spermatogenesis, the complete spermatogenesis cycle has not been efficiently achieved in vitro. This project proposes using advanced micro-extrusion technology to precisely fabricate microtubes with cellular composition and organization of the in vivo seminiferous tubules and surrounding interstitial tissue. For convenience, it is termed bio-fabricated seminiferous tubules (BioSTs). The hypothesis is that IVS could be efficiently achieved by establishing an in-vivo-like microenvironment. The proposed research is based on advanced cell culture technologies from the PI lab and decades of studies on spermatogenesis from the Co- PI’s lab. The PI has developed a new technology called AlgTubes for mammalian cell culture. AlgTubes culture cells in micro-scale alginate hydrogel tubes suspended in a cell culture medium. The microtubes provide cells with a physiology-relevant microenvironment, thus, significantly improving the culture efficiency such as viability, growth rate, and yield. AlgTubes have many similarities to the seminiferous tubules, such as their tubular shape, micro-scale diameter, and mass transport via diffusion through the tube wall. This project will repurpose and further develop AlgTubes to build BioSTs with three specific aims: to fabricate BioSTs, to characterize BioSTs including their architecture, blood-testis-barrier, spermatogonia stem cells maintenance, male hormone production, and in vitro spermatogenesis for three months, and to validate BioSTs by modeling findings from a previously studied mouse model with Premel1 knockout. BioSTs will be invaluable for advancing the knowledge of spermatogenesis and male infertility. It also can be an alternative to animal testing in high-throughput toxicity assays. This will reduce animal testing use and provide more accurate and relevant results, as the in-vitro sperm will be derived from human cells. Moreover, prepubertal boys undergoing gonadotoxic treatments are at risk for depleting their spermatogonial stem cells (SSCs). In-vitro-derived spermatozoa can provide a solution for those boys. Similarly, the technology will benefit the infertility treatment of non-obstructive azoospermia patients who cannot produce spermatozoa but still have SSCs.

项目总结