Isolation, characterization and transduction efficiency of ovine Muse cells for translational research in cardiac regeneration

CASTILLO, MARTHA GIOVANNA; BAUZÁ, MARÍA DEL ROSARIO; LOCATELLI, PAOLA; CROTTOGINI, ALBERTO; OLEA, FERNANDA DANIELA; CUNIBERTI, LUIS

La Plata

Congreso; Reunión anual de la Sociedad Argentina de Fisiología.; 2021

Introduction: Multilineage differentiating stress-enduring (Muse) cells are non-tumorigenic endogenous pluripotent-like stem cells identified as SSEA+ cells. However, their features in large mammals needed for translational cardiac regeneration researches have not been described.Objective: Since ovine models of cardiac diseases are being increasingly used, we aimed at isolating Muse cells from ovine adipose tissue and characterize them. In addition, we tested the transduction efficiency of a baculoviral vector to genetically modify these cells.Methods: Ovine Muse cells were isolated from abdominal adipose tissue under a stress procure and cultured in suspension for 5 days and then expanded in adherence until passage 4. Mesenchymal stem cells (ASC) were also isolated. Muse cells were labeled with an antibody against the embryonic marker SSEA3. Gene expression of pluripotency markers (Nanog and Oct4) and S1P receptor 2 (S1PR2) was measured by RTqPCR in Muse cells and ASC. Ovine Muse cells were then transduced at different multiplicity of infection (MOI) with a baculoviral vector expressing the green fluorescent protein (Bv.CMV-GFP). The optimal transduction efficiency (TE) was determined by flow cytometry. Data were analyzed using Student-test and expressed as mean±SD.Results: Muse cells were 99.81% positive for the SSEA3 compared with 14,60% in ASC. TE was MOI 100: 75,86%; 200: 84,30% and 400: 87,21%. Relative gene expression in Muse cells was for S1PR2 (2.7± 1.1 vs.1.0±0.2, p