Page 80 - Eshre 2018

Dissociated testicular cells cultured in vitro on a
3D matrix: is it a good option?
•Preparation of single-cell suspension of testicular tissue from immature mice
•In vitro culture in the soft agar or methylcellulose matrices, providing a 3D system, + gonadotrophins
Stukenborg et al 2009 Mol. Hum. Reprod.
•Germ cell progression and maturation: meiotic pachytene spematocytes at 3 weeks of culture, spermatozoa after 31 days of culture
•No proof of the fertilization potential of in vitro produced sperm
•Maintenance of testosterone production in vitro •Very low efficiency!!!
Human testicular organoids: an approach to reconstitute in vitro a testis-like organ
Baert et .2017 Stem cell reports
•Preparation of a natural scaffold formed by a decellularized testicular matrix obtained from different patients
•Isolation of primary testicular cells from human adult or pubertal patients
•Inoculation of testis cells onto the scaffold and in vitro culture for up to 4 weeks at the gas-liquid interphase
•Evaluation of the organoid organization by immunofluorescence analysis of different cell type specific markers
•Testosterone, inhibin and cytokine secretion
•Advantage: maintenance of cell identity/function •Limitation: loss of testicular cytoarhitecture
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