Methods of expanding human prepubertal spermatogonial stem cells
AssigneeNam Tran
InventorsNAM TRAN, JAMES SMITH
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Abstract
The invention comprises methods of identifying and isolating spermatagonial stem cells in testicular tissue samples by the use of markers unique to spermatagonial stem cells. The invention defines and characterizes a unique testicular cellular niche wherein spermatagonial stem cells reside. The invention also comprises methods of identifying and isolating multipotent testicular stromal cells, on which spermatagonial stem cells can be cultured. Cultured spermatagonial stem cells, derived from prepubertal males or cancer patients can be utilized to generate viable spermatozoon or may be subsequently transplanted back into the donor to restore fertility.
Claims12
1-23. (canceled)
A method of producing spermatogonial stem cells comprising,
- co-culturing of THY1+/SSEA4− cells isolated from a testicular tissue sample or their progeny with THY1−/SSEA4+ cells isolated from a testicular tissue sample;
- and
- allowing colonies of the THY1−/SSEA4+ cells to form and expand.
A method of instilling fertility in a male subject, comprising
- obtaining a testicular tissue sample from the male subject;
- digesting the testicular tissue sample to create a single cell suspension;
- isolating THY1−/SSEA4+ cells from the single cell suspension;
- co-culturing the THY1−/SSEA4+ cells with THY1+/SSEA4− cells derived from a testicular tissue sample;
- allowing colonies of the THY1−/SSEA4+ cells to form and expand;
- harvesting the THY1−/SSEA4+ cells and cryopreserving them;
- and
- transplanting the cryopreserved THY1−/SSEA4+ cells into the testes of the male subject.
A method of isolating spermatogonial stem cells, comprising
- obtaining a testicular tissue sample from a male subject;
- digesting the testicular tissue sample to create a single cell suspension;
- isolating THY1−/SSEA4+ cells from the single cell suspension.
Dependencies are inferred from the claim text.