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  • ISSN: 2379-0490
    Early Online
    Volume 4, Issue 1
    Research Article
    Tetsutaro Kikuchi and Tatsuya Shimizu*
    Abstract: Purification of skeletal muscle myoblasts from other cells such as fibroblasts is essential in basic research and cell-based regenerative medicine. In this report a simple non-serological method is proposed to purify desmin-positive myoblasts using Brefeldin A.
    Ordinary passaged human skeletal muscle-derived cells were cultured for 3 days with five exocytic pathway inhibitors (Brefeldin A, Monensin, AACOCF3, Exo-1 and Golgicide A). The myoblasts identified by desmin exhibited relatively superior growth compared to the other cells, both in 7 - 15 ng/mL of Brefeldin A and in 850 – 2500 ng/mL of Golgicide A. In the case of 10 ng/mL Brefeldin A, the cell numbers increased 6.7-fold in desmin-positive cells and 2.4-fold in desmin-negative cells while they increased 11.7-fold and 15.6-fold in absence of Brefeldin A, respectively. In addition, 7-day cultures in 10 ng/mL Brefeldin A of four independent cell strains suggested that moderate starting purities resulted in high purities over 90%, while extremely low starting purities (< 1%) did not support stable purifications. Lastly, a strain purified through a 7-day culture with 10 ng/mL Brefeldin A was subjected to myotube induction culture. Fluorescent imaging using a confocal microscope revealed their ability to form multinucleated myotubes and myofibers with sarcomeric structures.
    In conclusion, the purification of desmin-positive cells by culturing human skeletal muscle-derived cells with Brefeldin A provides a simple method to obtain purified myoblasts for use in basic research and cell-based regenerative medicine.
    Short Communication
    Aline Evangelista Aguiar, Mariana de Oliveira Silva, Bianca Cheregatti Longo, Maria do Carmo Gonçalves, Celso Aparecido Bertran*
    Scaffold porosity is an essential factor for tissue growth since it allows cell proliferation and vascularization. In this work, three different scaffolds were produced by easy methods. The first one was prepared using Bioglass 45S5 particles and H2O2 as a foaming agent. In the second, glass nanoparticles were arranged as porous structure by the self-assembling of cellulose nanocrystals. In the last, a polymeric xanthan/chitosan composite scaffold was prepared by swelling method. The results showed that the scaffolds presented pore morphology, wall thickness and size distribution that are dependent on the materials and preparation method used, and that they are suitable for a range of applications.
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