Peripheral blood mononuclear cells contribute to myogenesis in a 3D bioengineered system of bone marrow mesenchymal stem cells and myoblasts

Pasqualina Scala; Paola Manzo; Erwin Pavel Lamparelli; Joseph Lovecchio; Maria Camilla Ciardulli; Valentina Giudice; Carmine Selleri; Emanuele Giordano; Laura Rehak; Nicola Maffulli; Giovanna Della Porta

doi:10.3389/fbioe.2022.1075715

Peripheral blood mononuclear cells contribute to myogenesis in a 3D bioengineered system of bone marrow mesenchymal stem cells and myoblasts

Front Bioeng Biotechnol. 2023 Jan 10:10:1075715. doi: 10.3389/fbioe.2022.1075715. eCollection 2022.

Authors

Pasqualina Scala¹, Paola Manzo^{1

2}, Erwin Pavel Lamparelli¹, Joseph Lovecchio³, Maria Camilla Ciardulli¹, Valentina Giudice^{1

2}, Carmine Selleri^{1

2}, Emanuele Giordano³, Laura Rehak⁴, Nicola Maffulli^{1

5}, Giovanna Della Porta^{1

6}

Affiliations

¹ Department of Medicine, Surgery and Dentistry, University of Salerno, Baronissi, Italy.
² Hematology and Transplant Center, University Hospital "San Giovanni di Dio e Ruggi D'Aragona", Salerno, Italy.
³ Department of Electrical, Electronic and Information Engineering "Guglielmo Marconi" (DEI), University of Bologna, Bologna, Italy.
⁴ Athena Biomedical innovations, Florence, Italy.
⁵ Centre for Sports and Exercise Medicine, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, England.
⁶ Interdepartment Centre BIONAM, University of Salerno, Fisciano, Italy.

Abstract

In this work, a 3D environment obtained using fibrin scaffold and two cell populations, such as bone marrow-derived mesenchymal stem cells (BM-MSCs), and primary skeletal muscle cells (SkMs), was assembled. Peripheral blood mononuclear cells (PBMCs) fraction obtained after blood filtration with HemaTrate^® filter was then added to the 3D culture system to explore their influence on myogenesis. The best cell ratio into a 3D fibrin hydrogel was 1:1 (BM-MSCs plus SkMs:PBMCs) when cultured in a perfusion bioreactor; indeed, excellent viability and myogenic event induction were observed. Myogenic genes were significantly overexpressed when cultured with PBMCs, such as MyoD1 of 118-fold at day 14 and Desmin 6-fold at day 21. Desmin and Myosin Heavy Chain were also detected at protein level by immunostaining along the culture. Moreover, the presence of PBMCs in 3D culture induced a significant downregulation of pro-inflammatory cytokine gene expression, such as IL6. This smart biomimetic environment can be an excellent tool for investigation of cellular crosstalk and PBMC influence on myogenic processes.

Keywords: 3D Co-culture model; mesenchymal stem cells; myogenesis; peripheral blood mononuclear cells; tissue engineering.

Grants and funding

The present study was only partially funded by the Athena Srl and mainly funded by the University of Salerno. Collection, analysis, interpretation of data or the decision to submit it for publication. LR provided the methodology for PBMC fraction collection; she also provided a literature survey useful for the organization of the introduction section.