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The Key to Improving Your Cell Culture

Hypoxic cell culture has been shown to improve cell culture significantly, like cell functionality and yield. The CytoSMART™ System allows monitoring of cell cultures within defined incubator conditions such as in hypoxia. In a recent study, Henn and coworkers found that human Mesenchymal Stem Cell yields were higher when kept in optimal hypoxic conditions (using the Xvivo System X3 for cell incubation, BioSpherix), compared to cells that were interrupted for handling and inspection.


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Hypoxic Culture Conditions Mimic In Vivo Environment of Cells

The use of hypoxic, i.e. low oxygen, culture conditions for cell cultures finds more and more interest in research as a means to mimic the in vivo situation more closely. Whereas physiological in vivo oxygen concentrations can range from 1% to 15%, most cell cultures are maintained in room air oxygen conditions (~ 21% O2). However, the value of O2 control in addition to CO2 and temperature control has shown to e.g. heavily influence gene expression profiles and phenotypic changes. Hence hypoxic culture is increasingly applied in stem cell and cancer research, in toxicological studies as well as in clinical-grade stem cell production.

While hypoxic conditions can easily be established using specialized incubators, the monitoring of cell cultures under hypoxic conditions has been extremely restricted until now. An online-monitoring system that allows direct monitoring of the cell culture in the incubator would be the preferred option, which so far however, was not readily available to researchers. The CytoSMART™ System overcomes this issue in that it allows continuous online monitoring of your cell culture under hypoxic culture. With its small size (133 x 90 x 100 mm), the CytoSMART™ Device fits into virtually any specialized incubator. What is more, as the cell culture can be viewed via the CytoSMART™ Cloud Solution, the monitoring of hypoxic cultures becomes completely independent of the presence of the researcher in the lab.

 

Hypoxic Culture in Stem Cell Applications

Photo of hMSCs grown under hypoxia taken with the CytoSMART System

Image of human mesenchymal stem cells cultured under hypoxic conditions taken with the CytoSMART™ Lux 10X Device. (Image courtesy of A. Henn, BioSpherix)

Hypoxic culture conditions are thought to support stem cell performance in general. The use of physiologically relevant oxygen levels in stem cell culture is beneficial, for example, for the generation of induced pluripotent stem cells (iPSC) and in mesenchymal stem cell (MSC) culture.


iPSCs have the ability to self-renew and generate any cell type in the human body. Therefore, they have the potential to produce an infinite quantity of cells for different applications, such as regenerative medicine, disease modeling and drug development. Many efforts have been made to find ways to facilitate the reprogramming process as well as the maintenance of iPSCs. Culturing iPSCs in undisturbed hypoxic conditions has been found to be one of the players that can elicit significant improvements to stem cell culture.

 

Enhanced iPSC generation, reduced spontaneous differentiation, and enhanced clonality of human embryonic stem cells have been demonstrated to be a result of hypoxia-induced stabilization of transcription factors of the Hypoxia-inducible factor (HIF) family.

The use of human mesenchymal stem cells (hMSCs) as a possible therapeutic tool in regenerative medicine has been widely assessed. Poor growth kinetics, early senescence, and genetic instability during in vitro expansion are among others the major challenges for MSC-based regenerative therapies. By applying physiological oxygen levels to the cell culture, major improvements have been achieved, such as modulated angiogenic potential of MSCs, reduced senescence and increased proliferation levels1. Furthermore, the quantity and quality of MSCs secreted bioactive molecules (called MSC secretome) diverge under hypoxic conditions. The number of different proteins expressed in hMSCs cultured under hypoxic conditions is much higher (see graph below), and secreted proteins that are identical between hypoxic and room air oxygen conditions show significantly different expression levels. These findings support why the MSC secretome is being considered a critical element for therapeutic efficacy of hMSC.

 

 

 

 

Analysis of human umbilical cord Wharton Jelly-derived hMSCs conditioned medium (hWJ-MSC CM): More proteins were identified in the hWJ-MSC CM collected from hypoxic conditions (166 proteins) when compared to the normoxic hWJ-MSC CM (104 proteins), in which 81 were common to the two conditions1.



Hypoxic Culture in Cancer Research

Low O2 concentration, or hypoxia, often occurs in the center of solid tumors where the presence and formation of blood vessels is often abnormal or limited. Hence low oxygen conditions play a key role in regulating cancer cell metabolism and the deeper understanding of this correlation may advance our understanding of cancer formation and the development of potential new therapies. 

Application example of hypoxic cultures in cancer research:

Reaction of normoxic vs. hypoxic colorectal cancer cell lines against the cancer drug AW464 (NSC 706704; a novel benzothiazole substituted quinol compound) showed that hypoxic cells are more resistant to the drug. Comparisons to normal cells such as endothelial cells or fibroblasts (both from Lonza) revealed drug potency against endothelial cells but not fibroblasts, suggesting possible anti-angiogenic activity2.

 

Enhance the quality of your research now by applying hypoxic culture conditions. With the CytoSMART™ System your hypoxic culture is always only one click away. Don’t hesitate and request a quote for your personal CytoSMART™ System.

 

References:

1. Teixeira FG, Panchalingam KM, Anjo SI, Manadas B, Pereira R, Sousa N, António J. Salgado AJ, Behie LA. (2015) Stem Cell Res Ther, 6:133

2. Mukherjee A, Westwell AD, Bradshaw TD, Stevens MFG, Carmichael J, Martin SG. (2005) Br J Cancer. 92: 350  

 

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