Generation of Organized Porcine Testicular Organoids in Solubilized Hydrogels from Decellularized Extracellular Matrix
Commonly used acronym: Testicular organoids
Scope of the method
- Human health
- Basic Research
- Translational - Applied Research
- In vitro - Ex vivo
- Animal derived cells / tissues / organs
- artificial testis
- extracellular matrix
- immature testicular tissue
- spermatogonial stem cells
- 3D Cell culture
- fertility preservation
- fertility restoration
This method describes the generation of porcine testicular organoids using piglet testicular cells seeded in a testicular extracellular matrix (tECM) hydrogel. To generate the solublized tECM hydrogel, porcine immature testicular tissues (ITTs) were dissected in small fragments and decellularized in a 0.01% sodium dodecyl sulfate solution followed by agitation in a 1% Triton X-100 solution before being lyophilized and digested in a solution of HCl/pepsin. To generate the organoids, testicular cell suspensions were isolated from the porcine ITT and seeded into the hydrogel to form each organoid (500 000 cells per hydrogel). The generated organoids were then cultured in vitro and showed both an architecture and endocrine function that are similar to that found in the native organ in vivo.
- - Biosafety cabinet,
- - Laminar flow hood,
- - Culture incubator,
- - Nanodrop,
- - Rheometer,
- - Mass spectrometer,
- - Cryogenic freezer,
- - Bain Marie.
- Published in peer reviewed journal
Pros, cons & Future potential
This an open acess method that describes in detail how to create an artificial porcine testis using testicular cell suspensions seeded in an 'inhouse' produced hydrogel formed from decellularized extracellular matrix.
An optimal balance between cell removal and extracellular matrix preservation is the biggest challenge of the technique.
The impact of tECM of different stiffnesses on the outcome of testicular organoids culture still needs to be evaluated.
This method can be potentially applied to testis tissue from other species to produce testis organoids.
References, associated documents and other information
Development of a Cytocompatible Scaffold from Pig Immature Testicular Tissue Al…
Contact personMarc Kanbar
OrganisationsUniversité Catholique de Louvain (UCL)
Institut de Recherche Expérimentale et Clinique (IREC), Andrology Lab