Fertility and sterility · 2014
Eliminating acute lymphoblastic leukemia cells from human testicular cell cultures: a pilot study.
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Sadri-Ardekani Hooman, Homburg Christa H, van Capel Toni M M, van den Berg Henk, van der Veen Fulco, van der Schoot C Ellen, van Pelt Ans M M, Repping Sjoerd
Аннотация
OBJECTIVE: To study whether acute lymphoblastic leukemia (ALL) cells survive in a human testicular cell culture system.
DESIGN: Experimental laboratory study.
SETTING: Reproductive biology laboratory, academic medical center.
PATIENT(S): Acute lymphoblastic leukemia cells from three patients and testicular cells from three other patients.
INTERVENTION(S): Acute lymphoblastic leukemia cells were cultured alone or in combination with testicular cells, at various concentrations, in a system that has recently been developed to propagate human spermatogonial stem cells.
MAIN OUTCOME MEASURE(S): Viability of ALL and testicular cells during culture was evaluated by flow cytometry using markers for live/dead cells. Furthermore, the presence of ALL cells among testicular cells was determined by highly sensitive (1:10,000 to 1:100,000 cells) patient-specific antigen-receptor minimal residual disease polymerase chain reaction. The presence of spermatogonia at the end of culture was determined by reverse transcription-polymerase chain reaction for ZBTB16, UCHL1, and GPR125.
RESULT(S): The ALL cells cultured separately did not survive beyond 14 days of culture. When cultured together with testicular cells, even at extremely high initial concentrations (40% ALL cells), ALL cells were undetectable beyond 26 days of culture. Reverse transcription-polymerase chain reaction confirmed the presence of spermatogonia at the end of the culture period.
CONCLUSION(S): Our pilot study shows that the described testicular cell culture system not only allows for efficient propagation of spermatogonial stem cells but also eliminates contaminating ALL cells.
DESIGN: Experimental laboratory study.
SETTING: Reproductive biology laboratory, academic medical center.
PATIENT(S): Acute lymphoblastic leukemia cells from three patients and testicular cells from three other patients.
INTERVENTION(S): Acute lymphoblastic leukemia cells were cultured alone or in combination with testicular cells, at various concentrations, in a system that has recently been developed to propagate human spermatogonial stem cells.
MAIN OUTCOME MEASURE(S): Viability of ALL and testicular cells during culture was evaluated by flow cytometry using markers for live/dead cells. Furthermore, the presence of ALL cells among testicular cells was determined by highly sensitive (1:10,000 to 1:100,000 cells) patient-specific antigen-receptor minimal residual disease polymerase chain reaction. The presence of spermatogonia at the end of culture was determined by reverse transcription-polymerase chain reaction for ZBTB16, UCHL1, and GPR125.
RESULT(S): The ALL cells cultured separately did not survive beyond 14 days of culture. When cultured together with testicular cells, even at extremely high initial concentrations (40% ALL cells), ALL cells were undetectable beyond 26 days of culture. Reverse transcription-polymerase chain reaction confirmed the presence of spermatogonia at the end of the culture period.
CONCLUSION(S): Our pilot study shows that the described testicular cell culture system not only allows for efficient propagation of spermatogonial stem cells but also eliminates contaminating ALL cells.