Cultivation of mammalian cells in heat-sealable pouches that are permeable to carbon dioxide

Klebe, Robert J.; Lyn, Shauna; Magnuson, Victoria L.; Zardeneta, Gustavo

doi:10.1016/0014-4827(90)90176-b

Cited by 3 publications

(2 citation statements)

References 21 publications

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“…Significantly, we demonstrate that cells can proliferate and potentially differentiate on scaffolds within macrofluidic systems. Using our approach, we demonstrate a decrease in overall bioreactor prototyping and construction costs and potentially answer the need for scalable bioreactor solutions for research and industry [49][50][51]. This study addresses this gap by leveraging laser welding to integrate nylon bilayers, potentially enabling affordable single-use bioreactors for laboratory-scale and industrial use [52][53][54][55][56].…”

Section: Introductionmentioning

confidence: 99%

Cultivation of Bovine MSCs on Plant-Based Scaffolds in a macrofluidic Single-Use Bioreactor for Cultured Meat

Gome,

Chak,

Tawil

et al. 2024

Preprint

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One of the main challenges of cultured meat is reducing the costs of production, often described as scaling. The main components of the cultivation process include: cells, media, scaffolds and bioreactors. This work presents an alternative to current stainless steel or glass bioreactors and uses food-grade plant-based scaffolds. Bioreactors must be sterile and leak-free to maintain a proper environment for cell growth. Thermoplastic films are a popular material for constructing these fluidic systems, but traditional methods of welding these film are expensive and do not allow rapid prototyping resulting in high research and development costs for companies. In this study, a laser welding method was developed to join thermoplastic films in a way that prevents contamination and leaks, affording effective macro-fluidics fabrication. This technique was tested using polyethylene (PET) films and a laser cutter operating with settings calibrated for the material to be welded or cut. The laser welding method was found to produce strong, leak-free seals in PET films with minimal heat-induced damage to the films. The ability to design fluidics, chambers and ports of various size. Using this method afforded the incorporation of plant-based scaffolds from food-grade plants (Rice). The laser welding method developed in this study provides a reliable, contamination-free method for the rapid fabrication of fluidic systems for cell cultivation. This technique has the potential to be compatible with a vast range of cell types and scaffolds and to be widely adopted in tissue engineering for regenerative medicine and food applications such as cultured meat.

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Section: Introductionmentioning

confidence: 99%

Cultivation of Bovine MSCs on Plant-Based Scaffolds in a macrofluidic Single-Use Bioreactor for Cultured Meat

Gome,

Chak,

Tawil

et al. 2024

Preprint

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“…In a few cases, they even reached one year (Schmied et al 2000) and beyond (Potter and Demarse 2001). Most of the presented culture devices with or without perifusion capabilities were designed as stand-alone systems intended to replace standard culture vessels (Ince et al 1985;Bavister 1988;Forsythe and Coates 1988;Klebe et al 1990;Pentz and Horler 1992;Hing et al 2000;Leclerc et al 2004;Prokop et al 2004;Ho et al 2005;De Bartolo et al 2006;Petronis et al 2006).…”

Section: Introductionmentioning

confidence: 99%

Replica-moulded polydimethylsiloxane culture vessel lids attenuate osmotic drift in long-term cell cultures

et al. 2009

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An imbalance in medium osmolarity is a determinant that affects cell culture longevity. Even in humidified incubators, evaporation of water leads to a gradual increase in osmolarity over time. We present a simple replica-moulding strategy for producing self-sealing lids adaptable to standard, small-size cell-culture vessels. They are made of polydimethylsiloxane (PDMS), a flexible, transparent and biocompatible material, which is gas-permeable but largely impermeable to water. Keeping cell cultures in a humidified 5% CO2 incubator at 37 degrees C, medium osmolarity increased by +6.86 mosmol/kg/day in standard 35 mm Petri dishes, while PDMS lids attenuated its rise by a factor of four to changes of +1.72 mosmol/kg/ day. Depending on the lid membrane thickness,pH drifts at ambient CO2 levels were attenuated by a factor of 4 to 9. Comparative evaporation studies at temperatures below 60 degrees C yielded a 10-fold reduced water vapour flux of 1.75 g/day/ dm 2 through PDMS lids as compared with 18.69 g/day/dm 2 with conventional Petri dishes. Using such PDMS lids,about 2/3 of the cell cultures grew longer than 30 days in vitro. Among these,the average survival time was 69 days with the longest survival being 284 days under otherwise conventional cell culture conditions.

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Cultivation of Bovine Mesenchymal Stem Cells on Plant-Based Scaffolds in a Macrofluidic Single-Use Bioreactor for Cultured Meat

Gome,

Chak,

Tawil

et al. 2024

Foods

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Reducing production costs, known as scaling, is a significant obstacle in the advancement of cultivated meat. The cultivation process hinges on several key components, e.g., cells, media, scaffolds, and bioreactors. This study demonstrates an innovative approach, departing from traditional stainless steel or glass bioreactors, by integrating food-grade plant-based scaffolds and thermoplastic film bioreactors. While thermoplastic films are commonly used for constructing fluidic systems, conventional welding methods are cost-prohibitive and lack rapid prototyping capabilities, thus inflating research and development expenses. The developed laser welding technique facilitates contamination-free and leakproof sealing of polyethylene films, enabling the efficient fabrication of macrofluidic systems with various designs and dimensions. By incorporating food-grade plant-based scaffolds, such as rice seeded with bovine mesenchymal stem cells, into these bioreactors, this study demonstrates sterile cell proliferation on scaffolds within macrofluidic systems. This approach not only reduces bioreactor prototyping and construction costs but also addresses the need for scalable solutions in both research and industrial settings. Integrating single-use bioreactors with minimal shear forces and incorporating macro carriers such as puffed rice may further enhance biomass production in a scaled-out model. The use of food-grade plant-based scaffolds aligns with sustainable practices in tissue engineering and cultured-meat production, emphasizing its suitability for diverse applications.

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Cultivation of mammalian cells in heat-sealable pouches that are permeable to carbon dioxide

Cited by 3 publications

References 21 publications

Cultivation of Bovine MSCs on Plant-Based Scaffolds in a macrofluidic Single-Use Bioreactor for Cultured Meat

Cultivation of Bovine MSCs on Plant-Based Scaffolds in a macrofluidic Single-Use Bioreactor for Cultured Meat

Replica-moulded polydimethylsiloxane culture vessel lids attenuate osmotic drift in long-term cell cultures

Cultivation of Bovine Mesenchymal Stem Cells on Plant-Based Scaffolds in a Macrofluidic Single-Use Bioreactor for Cultured Meat

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