Optimal Culture Conditions for Constructing Durable Biografts for Repairing the Impaired Heart —Dynamic Cell Culture with Pre-seeding—

Abstract

Background: Tissue engineering with cell seeded biodegradable material has attracted attention as a novel means of treating the severely impaired heart. Here, we consider optimal preparation of a durable biograft using dynamic and static cultures. Methods: Vascular smooth muscle cells (VSMCs) derived from the rat aorta were seeded onto biodegradable material P (LA/CL) (poly-L-lactide-ε-caprolactone copolymer) and cultured as follows: a) Static culture (n = 11), b) dynamic culture (n = 12), c) 0 h pre-seeding (n = 12), d) 24 h pre-seeding (n = 5) and e) 1 week pre-seeding (n = 12). Dynamic culture: Cells were cultured in spinner flasks. Pre-seeding: Static cell seeding and culture before dynamic culture. Evaluation: The conditions of the P (LA/CL) in the five groups were evaluated as cell proliferation and by histological studies. Results: VSMCs proliferated both in and on the biodegradable materials. The quality of the dynamic culture cell with pre-seeding increased. Although the duration of pre-seeding exerted no significantly different effects, cell attachment and proliferation were widely scattered in the 0 h pre-seeding group, whereas cells proliferating on the front of the scaffold obstructed proliferation inside the biodegradable material in the 1 week pre-seeding group. Conclusions: Dynamic cell culture with 24 h pre-seeding is effective for constructing ideal biografts.

Canine Placenta Recellularized Using Yolk Sac Cells with Vascular Endothelial Growth Factor

“…There are many strategies adopted for the recellularization of biological scaffolds. Among them we can mention the use of bioreactors 36–38 and spinners flasks 39 , 40 or the immersion/agitation in detergent solutions.…”

Spherical rotary cell seeding system for production of small-caliber tissue-engineered blood vessels with complex geometry

“…The UV-C treatment of PETG is known to promote cell adherence by modifying the plastic’s carbonyl groups, which in turn augment the hydrophilic properties of the material 14 , 19 – 21 . In accordance with the Langmuir theory, the rotational movement of the described dynamic seeding system facilitates cell-scaffold interactions on the entire surface of the treated plastic mandrel, allowed for better cell adherence and distribution 7 , 8 , 22 . Indeed, the 360° random movement of this novel system allowed for cells to encounter all parts of the complex geometry scaffold during the seeding period.…”

“…One of the challenges in vascular tissue engineering is still, however, to improve cellular seeding, distribution, and organization to homogeneously incorporate cells onto a tubular structure. Consequently, dynamic cell seeding techniques have established themselves over simpler static approaches [7][8][9] . Furthermore, in a tri-dimensional (3D) environment, a uniformly monitored cell distribution is needed to foster homogeneous tissue remodeling and to avoid competition for nutrients in areas with higher cell densities [10][11][12][13] .…”

>b<Padronização dos processos de recelularização de >i<scaffolds>/i< biológicos provenientes de placentas caninas>/b<

“…Existem muitas estratégias adotadas para a recelularização de scaffolds biológicos. Entre eles podemos citar o uso de biorreatores (BONVILLAIN et al, 2013;KASUKONIS et al, 2016;ZHOU et al, 2016) e spinners flasks (YOKOMURO et al, 2011;TEIXEIRA et al, 2014) (ROH et al, 2007a(ROH et al, , 2007b escolhida com base no tipo e na porosidade do scaffold (MA, 2008). Além disso, o período relativamente longo de incubação de células no scaffold (7 dias) e, finalmente, a presença do VEGF e a natureza do biomaterial, podem ter favorecido o sucesso deste primeiro teste de recelularização.…”