Short-Term Culture of Human Neonatal Myofibroblasts Seeded Using a Novel Three-Dimensional Rotary Seeding Device

Abstract

A crucial factor in the tissue engineering of heart valves is an effective cell seeding with uniform cell distribution on biodegradable scaffolds to eventually form functional tissue constructs in vitro. In our laboratory, we developed a new cell-seeding device for optimal cell distribution for tissue-engineered heart valve constructs. In the present study, we developed a new cell-seeding device made of acrylic glass that is completely transparent (University Hospital Benjamin Franklin, Berlin, Germany). The polymeric heart valve scaffold is fixed in a small-volume, cylindrical cell-seeding chamber, and is surrounded by optimal cell suspension. The cell-seeding chamber is placed in a clear acrylic bowl so that it can be rotated in all directions to provide optimal cell distribution to all areas of the heart valve construct. We thus developed a highly isolated cell-seeding device that is driven by an independently developed rotating machine consisting of two independent motors (University Hospital Benjamin Franklin, Berlin, Germany). The whole system provides a high level of sterility and fits into a humidified incubator. Our newly developed cell-seeding device enables sterile conditions and optimal cell distribution for the controlled fabrication of autologous tissue-engineered heart valve constructs.

Cells, scaffolds and bioreactors for tissue-engineered heart valves: a journey from basic concepts to contemporary developmental innovations☆

“…Most bioreactors work at pulmonary pressure range (15-30 mm Hg), while the systemic one is often achieved as a differential value (DP) set at lower than physiological level. The most advanced bioreactor prototypes feature pneumatic pistons, which were more suitable for the reproduction of the in vivo heart physiology [22,24,74]. Because there is a highly nonlinear relation between the dynamic pulsatile nature and the pressure of the systemic blood flow (a direct consequence of the fluid and the viscoelastic properties of the tissue), bioreactor designers developed specific mechanisms for the independent fine-tuning of these parameters.…”

“…These data are related to both the mechanical stimuli to which cells are subjected, and to the mixing fluid movement responsible for the deeper gas and nutrient perfusion into engineered tissues [11,12]. Human bone marrow stem cells [13,14], endothelial progenitor cells (EPC) [15], amniotic fluid and chorionic villi mesenchymal stem cells (MSCs) [16,17], vascular umbilical cord elements [18,19], as well as from human and ovine vascular stroma [20][21][22][23][24][25][26][27] have been tested in experiments of acellular heart valve scaffold seeding. The full gamut of stem cells has demonstrated their plasticity, multipotency, and capacity for self-renewal by differentiating into the heart valve lineages (smooth muscle cells ((SMCs), fibroblasts, and myofibroblasts, that are also named vascular interstitial cells (VICs)) [28].…”

A New Bioreactor for the Development of Tissue-Engineered Heart Valves

“…The pressurized driving fluid thus allows a certain control of the flow rate waveform, depending on the complexity of the actuator device. A ventricular pump design was used in the bioreactors presented by Hoerstrup et al, 7 Sodian et al, 19 and Lueders et al 12 In all these studies, the driving fluid was compressed air provided by a commercial Harvard respirator. Although the reported waveforms were still somewhat primitive, they showed a significant improvement compared to those obtained with a peristaltic pump.…”

“…1 In the case of heart valve or arterial replacements, many specialists agree that biological substitutes are preferable to synthetic ones. 7,11,12,17 Limited life expectancy and post-surgical complications are regarded as the main disadvantages linked to mechanical and synthetic biomaterial substitutes. 5,7 Biological substitutes exhibit the advantageous potential of adaptation to the patient's cardiovascular environment characteristics and regeneration along the aging process.…”

Biomimetic 3D Tissue Models for Advanced High-Throughput Drug Screening

“…Current methods for engineering cardiac tissue on scaffolds require cells found in the native cardiac environment, such as endothelial cells, 33 smooth muscle cells, 43 myofibroblasts, 44 and fibroblasts. 45 These cells need to be able to self-assemble in the engineered ECM scaffolding in order to generate functional tissues.…”