Generation of Organized Porcine Testicular Organoids in Solubilized Hydrogels from Decellularized Extracellular Matrix

Commonly used acronym: Testicular organoids

Scope of the method

The Method relates to
  • Human health
The Method is situated in
  • Basic Research
  • Translational - Applied Research
Type of method
  • In vitro - Ex vivo
This method makes use of
  • Animal derived cells / tissues / organs
Species from which cells/tissues/organs are derived
Type of cells/tissues/organs


Method keywords
  • artificial testis
  • decellularization
  • extracellular matrix
  • immature testicular tissue
  • spermatogonial stem cells
  • testis
  • organoids
  • 3D Cell culture
Scientific area keywords
  • fertility preservation
  • cancer
  • boys
  • fertility restoration
Method description

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.

Lab equipment
  • - Biosafety cabinet,
  • - Laminar flow hood,
  • - Culture incubator,
  • - Nanodrop,
  • - Rheometer,
  • - Mass spectrometer,
  • - Cryogenic freezer,
  • - Bain Marie.
Method status
  • 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.

Future & Other applications

This method can be potentially applied to testis tissue from other species to produce testis organoids.

Contact person

Marc Kanbar


Université Catholique de Louvain (UCL)
Institut de Recherche Expérimentale et Clinique (IREC), Andrology Lab