Generation of spermatid-like cells from testicular biopsies obtained from prepubertal boys.

OBJECTIVE

To study whether functional human spermatids can be generated from spermatogonial stem cells obtained from testicular biopsies of prepubertal boys newly diagnosed with cancer using a three-dimensional (3D) human testicular organoid system.

DESIGN

Experimental laboratory study using a 3D human testicular organoid model incorporating Sertoli cells and endothelial cells to recreate the testes microenvironment.

SUBJECTS: Testicular biopsy specimens obtained from prepubertal boys newly diagnosed with cancer before initiation of gonadotoxic cancer therapy for fertility preservation.

EXPOSURE: Culture of prepubertal testicular tissue in a 3D human testicular organoid system with growth factors to support germ cell differentiation.

MAIN OUTCOME MEASURES: Generation of differentiated spermatids followed by verification of postmeiotic marker expression by immunofluorescence and assessment of functional competence through fertilization assays.

RESULTS

Germ cells differentiated into round spermatid-like cells within our testicular organoid system. Immunofluorescence confirmed expression of markers consistent with postmeiotic germ cell differentiation. Functional competence was demonstrated by successful fertilization assays using mouse oocytes, indicating that the spermatid-like cells generated ex vivo could activate oocytes.

CONCLUSION

Spermatid-like cells can be generated from prepubertal testicular tissue in a three-dimensional organoid culture system, supporting the potential feasibility of fertility preservation for boys undergoing gonadotoxic cancer treatments.