Tubular Perfusion System for the Long-Term Dynamic Culture of Human Mesenchymal Stem Cells

Abstract

In vitro culture techniques must be improved to increase the feasibility of cell-based tissue engineering strategies. To enhance nutrient transport we have developed a novel bioreactor, the tubular perfusion system (TPS), to culture human mesenchymal stem cells (hMSCs) in three-dimensional scaffolds. This system utilizes an elegant design to create a more effective environment for cell culture. In our design, hMSCs in the TPS bioreactor are encapsulated in alginate beads that are tightly packed in a tubular growth chamber. The medium is perfused by a peristaltic pump through the growth chamber and around the tightly packed scaffolds enhancing nutrient transfer while exposing the cells to shear stress. Results demonstrate that bioreactor culture supports early osteoblastic differentiation of hMSCs as shown by alkaline phosphatase gene expression. After 14 and 28 days of culture significant increases in the gene expression levels of osteocalcin, osteopontin, and bone morphogenetic protein-2 were observed with bioreactor culture, and expression of these markers was shown to increase with media flow rate. These results demonstrate the TPS bioreactor as an effective means to culture hMSCs and provide insight to the effect of long-term shear stresses on differentiating hMSCs.

In Vitro Biological Evaluation of 3-D Hydroxyapatite/Collagen (50/50 wt. (%)) Scaffolds

“…However, after this time, the number of cells decreased, and after 21 days, half of the cells had died, probably because of the culture conditions used (Figure 4b). Indeed, some authors have shown that to maintain viable cells inside 3-D scaffolds for long periods, it is necessary to force medium perfusion inside the scaffold using dynamic culture conditions [27][28][29] .…”

In Vitro Endothelialization of Biodegradable Vascular Grafts Via Endothelial Progenitor Cell Seeding and Maturation in a Tubular Perfusion System Bioreactor

“…31 Briefly, an L/S multichannel pump system (Cole Palmer) was used to drive flow (2 mL/min) through a tubing circuit. The flow rate was chosen based on previous data regarding EPC differentiation into EC-like cells.…”

“…We have previously demonstrated the successful application of the TPS bioreactor in the development and support of various tissue engineering constructs. 23,[29][30][31] In contrast to work showing the differentiation of EPCs in arterial shear stress environments, we chose to demonstrate cell seeding and differentiation in venous shear stress conditions given the lack of available TEVGs for venous conditions and their application in conditions such as congenital heart disease. Thus, it may be advantageous to understand the fate of EPCs within the venous system.…”

A flow perfusion bioreactor with controlled mechanical stimulation: Application in cartilage tissue engineering and beyond

“…The tubular perfusion bioreactors (TPBs) [49], are easily fabricated allowing cell construct encapsulatation in tightly packed spherical beads [50]. One study found hBMSCs encapsulated in alginate beads had 1.6 times more cells [49].…”

“…One study found hBMSCs encapsulated in alginate beads had 1.6 times more cells [49]. Others found hBMSCs secrete more GAGs in the dynamic environment of a TPB as evidenced by Alcian blue staining [51].…”