High throughput encapsulation of murine fibroblasts in alginate using the JetCutter technology

Schwinger, Christian; Koch, Sandra; Jahnz, Ulrich; Wittlich, Peter; Rainov, Nikolai G.; Kreßler, Jörg

doi:10.1080/02652040110105328

Cited by 36 publications

(15 citation statements)

References 20 publications

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“…46,47 Small (\1 mm) alginate beads are generated commonly by a vibrating nozzle, coaxial gas flow or cutting of a laminar jet by rotating blades (JetCutter). 48 The JetCutter, 49 the highest throughput laboratory-scale mammalian cell encapsulator described in the literature, would take [20 h to produce 10 L of alginate beads from a single nozzle. Lengthy and variable residence times of cells in the nonideal conditions of the gelling bath could be detrimental to cell health and lead to bead-to-bead variability.…”

Section: Potential Applicationsmentioning

confidence: 99%

A novel alginate hollow fiber bioreactor process for cellular therapy applications

Hoesli

Luu

Piret

2009

Biotechnology Progress

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Gel-matrix culture environments provide tissue engineering scaffolds and cues that guide cell differentiation. For many cellular therapy applications such as for the production of islet-like clusters to treat Type 1 diabetes, the need for large-scale production can be anticipated. The throughput of the commonly used nozzle-based devices for cell encapsulation is limited by the rate of droplet formation to approximately 0.5 L/h. This work describes a novel process for larger-scale batch immobilization of mammalian cells in alginate-filled hollow fiber bioreactors (AHFBRs). A methodology was developed whereby (1) alginate obstruction of the intra-capillary space medium flow was negligible, (2) extra-capillary alginate gelling was complete and (3) 83 +/- 4% of the cells seeded and immobilized were recovered from the bioreactor. Chinese hamster ovary (CHO) cells were used as a model aggregate-forming cell line that grew from mostly single cells to pancreatic islet-sized spheroids in 8 days of AHFBR culture. CHO cell growth and metabolic rates in the AHFBR were comparable to small-scale alginate slab controls. Then, the process was applied successfully to the culture of primary neonatal pancreatic porcine cells, without significant differences in cell viability compared with slab controls. As expected, alginate-immobilized culture in the AHFBR increased the insulin content of these cells compared with suspension culture. The AHFBR process could be refined by adding matrix components or adapted to other reversible gels and cell types, providing a practical means for gel-matrix assisted cultures for cellular therapy.

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Section: Potential Applicationsmentioning

confidence: 99%

A novel alginate hollow fiber bioreactor process for cellular therapy applications

Hoesli

Luu

Piret

2009

Biotechnology Progress

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“…Commonly used alginate bead generators rely on droplet formation by extrusion of an alginate and cell mixture through a nozzle and external gelation upon contact of the falling droplet with calcium or barium solution. However, the highest extrusion throughput allowing adequate droplet generation varies between $10 and $360 mL/h (Koch et al, 2003;Schwinger et al, 2002), requiring >1 day to generate 10 L of beads. In addition, the damage imparted by energy dissipation at the nozzle tip for high-viscosity alginate solutions at these flow rates has not been investigated (Shenkman et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Pancreatic cell immobilization in alginate beads produced by emulsion and internal gelation

Hoesli¹,

Raghuram²,

Kiang³

et al. 2010

Biotech & Bioengineering

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Alginate has been used to protect transplanted pancreatic islets from immune rejection and as a matrix to increase the insulin content of islet progenitor cells. The throughput of alginate bead generation by the standard extrusion and external gelation method is limited by the rate of droplet formation from nozzles. Alginate bead generation by emulsion and internal gelation is a scaleable alternative that has been used with biological molecules and microbial cells, but not mammalian cells. We describe the novel adaptation of this process to mammalian cell immobilization. After optimization, the emulsion process yielded 90 ± 2% mouse insulinoma 6 (MIN6) cell survival, similar to the extrusion process. The MIN6 cells expanded at the same rate in both bead types to form pseudo-islets with increased glucose stimulation index compared to cells in suspension. The emulsion process was suitable for primary pancreatic exocrine cell immobilization, leading to 67 ± 32 fold increased insulin expression after 10 days of immobilized culture. Due to the scaleability and broad availability of stirred mixers, the emulsion process represents an attractive option for laboratories that are not equipped with extrusion-based cell encapsulators, as well as for the production of immobilized or encapsulated cellular therapeutics on a clinical scale.

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“…The diameter of the beads produced by this technology ranges from 200 mm up to a few millimetres. With the JetCutter, Schwinger et al [20] successfully encapsulated murine fibroblasts into alginate/poly-L-lysine complexes, showing that encapsulated cells were able to survive in culture for extended periods of time with unchanged rates of proliferation and preserved morphology.…”

Section: Extrusionmentioning

confidence: 99%

Natural polymers for the microencapsulation of cells

Gasperini

Mano

Reis

2014

J. R. Soc. Interface.

531

413

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The encapsulation of living mammalian cells within a semi-permeable hydrogel matrix is an attractive procedure for many biomedical and biotechnological applications, such as xenotransplantation, maintenance of stem cell phenotype and bioprinting of three-dimensional scaffolds for tissue engineering and regenerative medicine. In this review, we focus on naturally derived polymers that can form hydrogels under mild conditions and that are thus capable of entrapping cells within controlled volumes. Our emphasis will be on polysaccharides and proteins, including agarose, alginate, carrageenan, chitosan, gellan gum, hyaluronic acid, collagen, elastin, gelatin, fibrin and silk fibroin. We also discuss the technologies commonly employed to encapsulate cells in these hydrogels, with particular attention on microencapsulation.

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High throughput encapsulation of murine fibroblasts in alginate using the JetCutter technology

Cited by 36 publications

References 20 publications

A novel alginate hollow fiber bioreactor process for cellular therapy applications

A novel alginate hollow fiber bioreactor process for cellular therapy applications

Pancreatic cell immobilization in alginate beads produced by emulsion and internal gelation

Natural polymers for the microencapsulation of cells

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