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Cytotechnology. 2018 Apr;70(2):651-664. doi: 10.1007/s10616-017-0166-4. Epub 2017 Nov 15.

Aligned ovine diaphragmatic myoblasts overexpressing human connexin-43 seeded on poly (L-lactic acid) scaffolds for potential use in cardiac regeneration.

Author information

1
Instituto de Medicina Traslacional, Transplante y Bioingeniería (IMETTYB), Universidad Favaloro-CONICET, Solís 453, C1078AAI, Buenos Aires, Argentina. sgimenez@favaloro.edu.ar.
2
Instituto de Medicina Traslacional, Transplante y Bioingeniería (IMETTYB), Universidad Favaloro-CONICET, Solís 453, C1078AAI, Buenos Aires, Argentina.
3
Instituto de Investigaciones en Ciencia y Tecnología de Materiales (INTEMA), Universidad Nacional de Mar del Plata-CONICET, Mar del Plata, Argentina.
4
Centro de Investigaciones Cardiológicas (CIC), Universidad Nacional de La Plata-CONICET, La Plata, Argentina.
5
Instituto de Investigaciones Biotecnológicas-Instituto Tecnológico de Chascomús (IIB-INTECH), Universidad Nacional de San Martín-CONICET, Chascomús, Argentina.

Abstract

Diaphragmatic myoblasts (DMs) are precursors of type-1 muscle cells displaying high exhaustion threshold on account that they contract and relax 20 times/min over a lifespan, making them potentially useful in cardiac regeneration strategies. Besides, it has been shown that biomaterials for stem cell delivery improve cell retention and viability in the target organ. In the present study, we aimed at developing a novel approach based on the use of poly (L-lactic acid) (PLLA) scaffolds seeded with DMs overexpressing connexin-43 (cx43), a gap junction protein that promotes inter-cell connectivity. DMs isolated from ovine diaphragm biopsies were characterized by immunohistochemistry and ability to differentiate into myotubes (MTs) and transduced with a lentiviral vector encoding cx43. After confirming cx43 expression (RT-qPCR and Western blot) and its effect on inter-cell connectivity (fluorescence recovery after photobleaching), DMs were grown on fiber-aligned or random PLLA scaffolds. DMs were successfully isolated and characterized. Cx43 mRNA and protein were overexpressed and favored inter-cell connectivity. Alignment of the scaffold fibers not only aligned but also elongated the cells, increasing the contact surface between them. This novel approach is feasible and combines the advantages of bioresorbable scaffolds as delivery method and a cell type that on account of its features may be suitable for cardiac regeneration. Future studies on animal models of myocardial infarction are needed to establish its usefulness on scar reduction and cardiac function.

KEYWORDS:

Absorbable implants; Connexin-43; Diaphragm; Fluorescence recovery after photobleaching; Myoblasts; Poly (L-lactic acid); Sheep

PMID:
29143226
PMCID:
PMC5851959
[Available on 2019-04-01]
DOI:
10.1007/s10616-017-0166-4

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