F127 (OR P407) POLOXAMER AS A PROMISING GELLING AGENT IN SERTOLI CELL DELIVERY FOR CELL THERAPY OF THE TESTIS

JACOBO P.; SOBARZO ALVAREZ, CM.; PEREZ, CV.; AMARILLA, MS.; LEILANE, G.; BERTINETTI, B.; FRUNGIERI, M.; GLISONI, RJ.; THEAS, S.

Mar del Plata

Congreso; S A I C . S A F E . S A B . SAP 2 0 1 9 Participan AACYTAL . NANOMED-AR . HCS; 2019

SAIC-NANOMED-ar

Sertoli cells (SCs) play an important role in creating an immune-privileged environment and supporting spermatogenesis. Their ability to survive transplantation long term without immunosuppression leads to the notion that SCs may be a tool for cell therapy of many chronic inflammatory diseases. Infertility has become a major health issue in the world, 40% due to male factor. There is no specific treatment to cure infertile patients; overall therapies try to bypass the cause using artificial reproductive technology. We hypothesize SC transplantation would be assessed as a practical approach for recovering spermatogenesis in azoospermic male with chronic inflammation. The trophic potential of SCs has been challenged by transplanting them in the testis of rats devoided of spermatogenesis by chemical treatment. However, experimental protocols employing a high number of SCs which disperse all around the testis had a very limited success. The aim of our work was to improve SC transfer protocols in order to increase cell density in specific areas of testis allowing diffusion of their secreted factors. We use biocompatible linear tri-block copolymer (Pluronic® F127) as cell delivery hydrogel and the SC line TM4. F127 was dispersed in saline to obtain 16-22% (w/v) dispersions and placed at 34°C. F127 preparation (22%) turned into hydrogel in less than 1 min. TM4 cells dispersed in 22% F127 kept viable up to 24 h at 34°C (Trypan blue method). 2-4x106 TM4 cells labeled with carboxyfluorescein-succinimidyl ester (CFSE) dispersed in 22% F127 or saline (100 µL) were injected ex-vivo in the testis from adult rats. After 7 min testes were frozen or fixed (PFA) for histology evaluation and fluorescence microscopy. Results showed that F127 increased cell density near injection sites limiting their dispersion vs cells transferred in saline. We propose that F127 is a useful copolymer matrix for cell therapy as it would allow secreted soluble factors reach therapeutic levels minimizing cell number requirement.