Portable, low‐cost hypoxia chamber for simulating hypoxic environments: Development, characterization and applications

Saxena, Shivanjali; Agrawal, Ishan; Singh, Pratibha; Jha, Sushmita

doi:10.1002/mds3.10064

Cited by 10 publications

(8 citation statements)

References 16 publications

(17 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Therefore, it is possible to selectively apply and eliminate only the wells in which the cells are cultured in pressure and low-oxygen environments. To change the existing cell culture environment from normoxia (20% O 2 concentration) [67] to hypoxia (1% O 2 concentration)[68], a wait of more than 2 h is required to stabilize the gas in the cell incubator [69] or to prepare a separate hypoxic incubator for replacement [70,71]. In addition, to apply pressure to the cells, a nancially burdensome hydrostatic pressure equipment [72,73] or a mechanical loading device [74,75] are required.…”

Section: Discussionmentioning

confidence: 99%

Biomimetic hydrogel blanket for conserving and recovering intrinsic cell properties

Seo

et al. 2022

Preprint

View full text Add to dashboard Cite

Background: Cells in the human body experience different growth environments and conditions, such as pressure and oxygen concentrations, depending on the type and location of the tissue. For this reason, a culture device that emulates the environment inside the body to study cells outside the body is required. Method: A blanket-type cell culture device (Direct Contact Pressing: DCP) was fabricated with an alginate-based hydrogel. Changes in cell morphology due to DCP pressure were observed with a phase-contrast microscope. The modification of oxygen permeability and pressure according to the hydrogel concentration of DCP was analyzed. To compare the effects of DCP with normal or artificial hypoxic culture, groups were divided into normal culture (Normal), use of DCP culture device (Pressing), and artificial hypoxia environment (Hypoxia). Changes in phenotype, genes, and GAG amounts according to each environment were evaluated. In addition, through the results of culture environment-gene and cell morphology analysis, the mechanism of each culture environment on the conserving chondrocytes intrinsic properties was suggested.Results: When human primary chondrocytes—which require pressure and a low-oxygen environment during culture to maintain their innate properties—were cultured using the hydrogel blanket, the original shapes and properties of chondrocytes were maintained, and recovery of intrinsic properties was observed even in aged cells which lost their original cell properties. Conclusion: A direct contact pressing culture method with the use of a biomimetic hydrogel blanket provides cells with adjustable physical pressure and a low-oxygen environment. Through this technique could maintain the original cellular phenotypes and intrinsic properties of human primary chondrocytes.

show abstract

Section: Discussionmentioning

confidence: 99%

Biomimetic hydrogel blanket for conserving and recovering intrinsic cell properties

Seo

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…Therefore, it is possible to selectively apply and eliminate only the wells in which the cells are cultured in pressure and low-oxygen environments. To change the existing cell culture environment from normoxia (20% O 2 concentration) [62] to hypoxia (1% O 2 concentration) [63], a wait of more than 2 h is required to stabilize the gas in the cell incubator [64] or to prepare a separate hypoxic incubator for replacement [65,66]. In addition, to apply pressure to the cells, financially burdensome hydrostatic pressure equipment [67,68] or mechanical loading device [20,69] are required.…”

Section: Discussionmentioning

confidence: 99%

Biomimetic hydrogel blanket for conserving and recovering intrinsic cell properties

Seo

et al. 2022

Biomater Res

View full text Add to dashboard Cite

Background Cells in the human body experience different growth environments and conditions, such as compressive pressure and oxygen concentrations, depending on the type and location of the tissue. Thus, a culture device that emulates the environment inside the body is required to study cells outside the body. Methods A blanket-type cell culture device (Direct Contact Pressing: DCP) was fabricated with an alginate-based hydrogel. Changes in cell morphology due to DCP pressure were observed using a phase contrast microscope. The changes in the oxygen permeability and pressure according to the hydrogel concentration of DCP were analyzed. To compare the effects of DCP with normal or artificial hypoxic cultures, cells were divided based on the culture technique: normal culture, DCP culture device, and artificial hypoxic environment. Changes in phenotype, genes, and glycosaminoglycan amounts according to each environment were evaluated. Based on this, the mechanism of each culture environment on the intrinsic properties of conserving chondrocytes was suggested. Results Chondrocytes live under pressure from the surrounding collagen tissue and experience a hypoxic environment because collagen inhibits oxygen permeability. By culturing the chondrocytes in a DCP environment, the capability of DCP to produce a low-oxygen and physical pressure environment was verified. When human primary chondrocytes, which require pressure and a low-oxygen environment during culture to maintain their innate properties, were cultured using the hydrogel blanket, the original shapes and properties of the chondrocytes were maintained. The intrinsic properties could be recovered even in aged cells that had lost their original cell properties. Conclusions A DCP culture method using a biomimetic hydrogel blanket provides cells with an adjustable physical pressure and a low-oxygen environment. Through this technique, we could maintain the original cellular phenotypes and intrinsic properties of human primary chondrocytes. The results of this study can be applied to other cells that require special pressure and oxygen concentration control to maintain their intrinsic properties. Additionally, this technique has the potential to be applied to the re-differentiation of cells that have lost their original properties.

show abstract

“…However, these systems may be cost-prohibitive and are challenging to customize for specific laboratory needs. To address this, several low-cost hypoxia chamber systems have been previously developed [37] , [38] , [39] . These systems were designed with ease-of-use, portability, and low-cost materials in mind.…”

Section: Hardware In Contextmentioning

confidence: 99%

“…In this system oxygen levels are controlled through gas displacement using N 2 , and a separate connection to a CO 2 tank, reducing the need for pre-mixed gases. Gas displacement involves inputting a gas (e.g., N 2 ) into the chamber, while having an exhaust port to displace the preexisting gas within the chamber, causing the overall O 2 concentration to decrease [39] . Thus, this low-cost and easy to reproduce hypoxia chamber can be used to explore how physiological oxygen levels impact cells and tissues in culture to advance basic science as well as tissue engineering and regenerative medicine.…”

Section: Hardware In Contextmentioning

confidence: 99%

Low-cost, open-source cell culture chamber for regulating physiologic oxygen levels

et al. 2022

View full text Add to dashboard Cite

The physiological oxygen levels for several mammalian cell types in vivo are considered to be hypoxic (low oxygen tension), but the vast majority of in vitro mammalian cell culture is conducted at atmospheric oxygen levels of around 21%. In order to understand the impact of low oxygen environments on cells, oxygen levels need to be regulated during in vitro culture. Two common methods for simulating a hypoxic environment are through the regulation of gas composition or chemical induction. Chemically mimicking hypoxia can have adverse effects such as reducing cell viability, making oxygen regulation in cell culture chambers crucial for long-term culture. However, oxygen-regulating cell culture incubators and commercial hypoxia chambers may not always be a viable option due to cost and limited customization. Other low-cost chambers have been developed, but they tend to lack control systems or are fairly small scale. Thus, the objective of this project was to design and develop a low-cost, open-source, controllable, and reproducible hypoxia chamber that can fit inside a standard cell culture incubator. This design allows for the control of O 2 between 1 and 21%, while maintaining CO 2 levels at 5%, as well as monitoring of temperature, pressure, and relative humidity. Testing showed our hypoxia chamber was able to maintain CO 2 levels at 5% and hypoxic O 2 levels at 1% and 5% for long-term cell culture. This simple and easy-to-manufacture design uses off the shelf components, and the total material cost was $832.47 (USD).

show abstract

Portable, low‐cost hypoxia chamber for simulating hypoxic environments: Development, characterization and applications

Cited by 10 publications

References 16 publications

Biomimetic hydrogel blanket for conserving and recovering intrinsic cell properties

Biomimetic hydrogel blanket for conserving and recovering intrinsic cell properties

Biomimetic hydrogel blanket for conserving and recovering intrinsic cell properties

Low-cost, open-source cell culture chamber for regulating physiologic oxygen levels

Contact Info

Product

Resources

About