Modelling human myoblasts survival upon xenotransplantation into immunodeficient mouse muscle

Abstract

Cell transplantation has been challenged in several clinical indications of genetic or acquired muscular diseases, but therapeutic success were mitigated. To understand and improve the yields of tissue regeneration, we aimed at modelling the fate of CD56-positive human myoblasts after transplantation. Using immunodeficient severe combined immunodeficiency (SCID) mice as recipients, we assessed the survival, integration and satellite cell niche occupancy of human myoblasts by a triple immunohistochemical labelling of laminin, dystrophin and human lamin A/C. The counts were integrated into a classical mathematical decline equation. After injection, human cells were essentially located in the endomysium, then they disappeared progressively from D0 to D28. The final number of integrated human nuclei was grossly determined at D2 after injection, suggesting that no more efficient fusion between donor myoblasts and host fibers occurs after the resolution of the local damages created by needle insertion. Almost 1% of implanted human cells occupied a satellite-like cell niche. Our mathematical model validated by histological counting provided a reliable quantitative estimate of human myoblast survival and/or incorporation into SCID muscle fibers. Informations brought by histological labelling and this mathematical model are complementary.

Combined methods to evaluate human cells in muscle xenografts

“…Analysis of the qPCR data showed a progressive diminution of the number of human myoblasts from 5d after grafting when compared to the amount found at injection (0d) (Fig 1D). This is in accordance with what is found in our and other previous works with human cells [12,16,17]: human myoblasts persist better than murine cells in mouse muscles and the massive loss of murine cells observed during the first days after injection [18] is not confirmed with human cells, when they are injected in an immunodeficient mouse, although a loss of cells is gradually observed at later time points (Fig 1D). More importantly, immediately after injection, human myoblasts remain confined in an enriched laminin “pocket” around the injection site [12].…”

Sıçanlarda immünsupresyonsuz xenotransplantasyon uygulamasının etkililiği

“…Bu yöntemlerin en radikal uygulaması, xenotransplantasyondur. Xenotransplantasyon uygulamaları başta domuz kullanılarak gerçekleştirilen kalp kapakçığı ve pankreas adacık hücre nakilleri olmak üzere 18 , kombine ağır immün yetmezliği (SCID) olan farelere insan myositlerinin nakli 19 , insan enterik sinir sistemini çalışmak amacıyla farelerin abdomenlerine uygulanan nakil işlemi 20 ya da insan kemik iliği hücrelerinin fonksiyonlarını araştırmak amacıyla kullanılan fare modelleri 21 gibi pek çok alanda uygulama imkanı bulmuştur. Ancak bu uygulama alanları arasında literatürde paratiroid hücrelerine spesifik olan çalışmalar oldukça sınırlı sayıdadır.…”