Microencapsulated Sertoli cells sustain myoblast proliferation without affecting the myogenic potential. In vitro data

Sara Chiappalupi; Laura Salvadori; Francesca Mancuso; Iva Arato; Mario Calvitti; Francesca Riuzzi; Riccardo Calafiore; Giovanni Luca; Guglielmo Sorci

doi:10.1016/j.dib.2021.107744

Microencapsulated Sertoli cells sustain myoblast proliferation without affecting the myogenic potential. In vitro data

Data Brief. 2021 Dec 23:40:107744. doi: 10.1016/j.dib.2021.107744. eCollection 2022 Feb.

Authors

Sara Chiappalupi^{1

2

3}, Laura Salvadori^{4

2

3}, Francesca Mancuso¹, Iva Arato¹, Mario Calvitti¹, Francesca Riuzzi^{1

2

3}, Riccardo Calafiore^{1

5}, Giovanni Luca^{1

5

6}, Guglielmo Sorci^{1

2

5

6

3}

Affiliations

¹ Department of Medicine and Surgery, University of Perugia, Perugia 06132, Italy.
² Interuniversity Institute of Myology (IIM), Perugia 06132, Italy.
³ Consorzio Interuniversitario Biotecnologie (CIB), Trieste 34127, Italy.
⁴ Department of Translational Medicine, University of Piemonte Orientale, Novara 28100, Italy.
⁵ Centro Biotecnologico Internazionale di Ricerca Traslazionale ad indirizzo Endocrino, Metabolico ed Embrio-Riproduttivo (CIRTEMER), Perugia 06132, Italy.
⁶ Centro Universitario di Ricerca sulla Genomica Funzionale (CURGeF), Perugia 06132, Italy.

Abstract

Sertoli cells (SeC) isolated from porcine testes have shown direct effects on muscle precursor cells sustaining C2C12 myoblasts proliferation and inhibiting oxidative stress and apoptosis in the early phase of the differentiation process, and stimulating myoblast fusion into myotubes and the expression of markers of myogenic differentiation in the late phase. This suggested that the cocktail of factors secreted by SeC stimulates proliferation in myoblasts without weakening their myogenic potential resulting in the formation of the critical myoblast amount necessary to rebuild the required muscle mass upon a damage. Here, we show that co-culturing C2C12 myoblasts with high doses of SeC microencapsulated in clinical grade alginate-based microcapsules (MC-SeC) for three days in differentiation medium (DM) translates into increased cell numbers and almost absence of myotube formation. However, after removal of MC-SeC, an intense fusion activity into myotubes was observed culminating in a fusion index similar to that of control after additional three days of culture in DM. These data definitely demonstrate that SeC-derived factors preserve the myogenic potential while sustaining cell proliferation in C2C12 myoblasts.

Keywords: ALG, sodium alginate; AMH, anti-Müllerian hormone; ASMA, alpha-smooth muscle actin; BSA, bovine serum albumin; DAPI, 4′,6-diamidino-2-phenylindole; DM, differentiation medium; E-MC, empty microcapsules; EB, ethidium bromide; EDTA, ethylene-diamine tetra-acetic acid; FBS, foetal bovine serum; FDA, fluorescein diacetate; Fusion index; HBSS, Hanks’ balanced salt solution; HG-DMEM, high-glucose Dulbecco’s modified Eagle’s medium; HS, horse serum; INSL3, insulin-like3; ITS, insulin-transferrin-selenium; MC-SeC, microencapsulated Sertoli cells; Microencapsulation; Myoblast; Myoblast proliferation; Myogenic potential; P/S, penicillin/streptomycin; PFA, paraformaldehyde; PGP9.5, protein gene product 9.5; SeC, Sertoli cells; Sertoli cell; TRIS, tris(hydroxymethyl)aminomethane.