Development of Postmeiotic Cells In Vitro from Spermatogonial Cells of Prepubertal Cancer Patients

Abstract

Aggressive chemotherapy in childhood often results in testicular damage and consequently jeopardizes future fertility. The presence of spermatogonial cells (SPGCs) in the testes of prepubertal cancer patient boys (PCPBs) can be used to develop future strategies for male fertility preservation. In the present study, we examined the presence of SPGCs in testes of chemotherapy-treated PCPBs and their ability to develop spermatogenesis in vitro using a three-dimensional culture system. Seven testicular biopsies were obtained from chemotherapy-treated PCPBs and one from a patient with β-thalassemia major. Isolated testicular cells were cultured in a methylcellulose culture system (MCS)-containing StemPro enriched with growth factors for 5-15 weeks. The presence of premeiotic, meiotic, and postmeiotic cells was examined by immunofluorescence staining and/or reverse transcription-polymerase chain reaction (RT-PCR) analysis. We observed SPGCs in the examined testicular biopsies. Isolated testicular cells cultured in MCS developed into colonies and contained premeiotic, meiotic, and postmeiotic cells. Furthermore, we identified sperm-like cells that had developed from testicular cells of a PCPB. Our results demonstrate, for the first time, the presence of biologically active SPGCs in testicular biopsies of chemotherapy-treated PCPBs and their capacity to develop in vitro to different stages of spermatogenesis, including the generation of sperm-like cells. This study may open the way for new therapeutic strategies for fertility preservation of PCPBs and for azoospermic patients.

Testicular Function of Childhood Cancer Survivors: Who Is Worse?

“…It is not yet certain if such cryopreserved tissue can be successfully used later to restore fertility in humans. However, the generation of sperm-like cells after 3D culture of isolated spermatogonial cells obtained from testis biopsies taken from pre-pubertal boys undergoing chemotherapy treatment has been recently described [21]. In a murine model, complete spermatogenesis was achieved through the in vitro culture of spermatogonial germ cells; the obtained mature spermatozoa have then been used to produce viable embryos through in vitro fertilization/ intracytoplasmic sperm injection (IVF/ICSI) [22].…”

Exposure to Chemotherapy During Childhood or Adulthood and Consequences on Spermatogenesis and Male Fertility

“…Today, only the organotypic culture allows to produce mature spermatozoa and generate a healthy offspring by intracytoplasmic sperm injection (ICSI) in mice [158,161]. In humans, studies are still limited and demonstrated a maturation arrest at the post-meiotic stage after 3D culture and organotypic culture from prepubertal and pubertal testicular tissue of cancer patient who has or has not received gonadotoxic treatment [162][163][164][165]. These data may pave the way for future studies aimed to optimise the conditions for in vitro maturation of frozen-thawed testicular tissue for future use in fertility preservation strategies.…”

Stem cell-based therapies for fertility preservation in males: Current status and future prospects

“…Compared with 2D culture, 3D culture can meet the requirement of recapitulating the natural physical structure of the human body and the microenvironment with its network of cell-cell interactions[ 112 ]. In mammals, 3D culture models have been more effective than 2D culture models[ 113 , 114 ]. In 1954, Trowell et al [ 115 ] first immersed semisolid-supported tissue fragments in culture medium to balance nutrient transport with effective gas exchange.…”