From Tissue Physoxia to Cancer Hypoxia, Cost-Effective Methods to Study Tissue-Specific O<sub>2</sub> Levels in Cellular Biology
The human body is endowed with an extraordinary ability to maintain different oxygen levels in various tissues and organs. The maintenance of physiological levels of oxygen is known as physoxia. The development of hypoxic conditions plays an important role in the biology of several pathologies, incl...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-05-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/10/5633 |
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| author | Carlos H. V. Nascimento-Filho Alexandra T. Glinos Yeejin Jang Eny M. Goloni-Bertollo Rogerio M. Castilho Cristiane H. Squarize |
| author_facet | Carlos H. V. Nascimento-Filho Alexandra T. Glinos Yeejin Jang Eny M. Goloni-Bertollo Rogerio M. Castilho Cristiane H. Squarize |
| author_sort | Carlos H. V. Nascimento-Filho |
| collection | DOAJ |
| description | The human body is endowed with an extraordinary ability to maintain different oxygen levels in various tissues and organs. The maintenance of physiological levels of oxygen is known as physoxia. The development of hypoxic conditions plays an important role in the biology of several pathologies, including cancer. In vitro studies using normal and neoplastic cells require that culture conditions be carried out under appropriate oxygen levels, either physoxic or hypoxic conditions. Such requirements are difficult to widely implement in laboratory practice, mainly due to the high costs of specialized equipment. In this work, we present and characterize a cost-effective method to culture cells under a range of oxygen levels using deoxidizing pouches. Our results show that physoxic and hypoxic levels using deoxidizing absorbers can be achieved either by implementing a gradual change in oxygen levels or by a regimen of acute depletion of oxygen. This approach triggers the activation of an epithelial-mesenchymal transition in cancer cells while stimulating the expression of HIF-1α. Culturing cancer cells with deoxidizing agent pouches revealed PI3K oncogenic pathway exacerbations compared to tumor cells growing under atmospheric levels of oxygen. Similar to the PI3K signaling disturbance, we also observed augmented oxidative stress and superoxide levels and increased cell cycle arrest. Most interestingly, the culture of cancer cells under hypoxia resulted in the accumulation of cancer stem cells in a time-dependent manner. Overall, we present an attractive, cost-effective method of culturing cells under appropriate physoxic or hypoxic conditions that is easily implementable in any wet laboratory equipped with cell culture tools. |
| first_indexed | 2024-03-10T03:43:49Z |
| format | Article |
| id | doaj.art-c3b338a60bba46b2a4d9adfdf6a57204 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T03:43:49Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-c3b338a60bba46b2a4d9adfdf6a572042023-11-23T11:26:02ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-05-012310563310.3390/ijms23105633From Tissue Physoxia to Cancer Hypoxia, Cost-Effective Methods to Study Tissue-Specific O<sub>2</sub> Levels in Cellular BiologyCarlos H. V. Nascimento-Filho0Alexandra T. Glinos1Yeejin Jang2Eny M. Goloni-Bertollo3Rogerio M. Castilho4Cristiane H. Squarize5Laboratory of Epithelial Biology, Department of Periodontics and Oral Medicine, University of Michigan School of Dentistry, Ann Arbor, MI 48109-1078, USALaboratory of Epithelial Biology, Department of Periodontics and Oral Medicine, University of Michigan School of Dentistry, Ann Arbor, MI 48109-1078, USALaboratory of Epithelial Biology, Department of Periodontics and Oral Medicine, University of Michigan School of Dentistry, Ann Arbor, MI 48109-1078, USAGenetics and Molecular Biology Research Unit (UPGEM), Department of Molecular Biology, School of Medicine of São José do Rio Preto, São José do Rio Preto 15090-000, SP, BrazilLaboratory of Epithelial Biology, Department of Periodontics and Oral Medicine, University of Michigan School of Dentistry, Ann Arbor, MI 48109-1078, USALaboratory of Epithelial Biology, Department of Periodontics and Oral Medicine, University of Michigan School of Dentistry, Ann Arbor, MI 48109-1078, USAThe human body is endowed with an extraordinary ability to maintain different oxygen levels in various tissues and organs. The maintenance of physiological levels of oxygen is known as physoxia. The development of hypoxic conditions plays an important role in the biology of several pathologies, including cancer. In vitro studies using normal and neoplastic cells require that culture conditions be carried out under appropriate oxygen levels, either physoxic or hypoxic conditions. Such requirements are difficult to widely implement in laboratory practice, mainly due to the high costs of specialized equipment. In this work, we present and characterize a cost-effective method to culture cells under a range of oxygen levels using deoxidizing pouches. Our results show that physoxic and hypoxic levels using deoxidizing absorbers can be achieved either by implementing a gradual change in oxygen levels or by a regimen of acute depletion of oxygen. This approach triggers the activation of an epithelial-mesenchymal transition in cancer cells while stimulating the expression of HIF-1α. Culturing cancer cells with deoxidizing agent pouches revealed PI3K oncogenic pathway exacerbations compared to tumor cells growing under atmospheric levels of oxygen. Similar to the PI3K signaling disturbance, we also observed augmented oxidative stress and superoxide levels and increased cell cycle arrest. Most interestingly, the culture of cancer cells under hypoxia resulted in the accumulation of cancer stem cells in a time-dependent manner. Overall, we present an attractive, cost-effective method of culturing cells under appropriate physoxic or hypoxic conditions that is easily implementable in any wet laboratory equipped with cell culture tools.https://www.mdpi.com/1422-0067/23/10/5633hypoxia chamberhypoxiaepithelial-mesenchymal transitioncancer stem cell |
| spellingShingle | Carlos H. V. Nascimento-Filho Alexandra T. Glinos Yeejin Jang Eny M. Goloni-Bertollo Rogerio M. Castilho Cristiane H. Squarize From Tissue Physoxia to Cancer Hypoxia, Cost-Effective Methods to Study Tissue-Specific O<sub>2</sub> Levels in Cellular Biology International Journal of Molecular Sciences hypoxia chamber hypoxia epithelial-mesenchymal transition cancer stem cell |
| title | From Tissue Physoxia to Cancer Hypoxia, Cost-Effective Methods to Study Tissue-Specific O<sub>2</sub> Levels in Cellular Biology |
| title_full | From Tissue Physoxia to Cancer Hypoxia, Cost-Effective Methods to Study Tissue-Specific O<sub>2</sub> Levels in Cellular Biology |
| title_fullStr | From Tissue Physoxia to Cancer Hypoxia, Cost-Effective Methods to Study Tissue-Specific O<sub>2</sub> Levels in Cellular Biology |
| title_full_unstemmed | From Tissue Physoxia to Cancer Hypoxia, Cost-Effective Methods to Study Tissue-Specific O<sub>2</sub> Levels in Cellular Biology |
| title_short | From Tissue Physoxia to Cancer Hypoxia, Cost-Effective Methods to Study Tissue-Specific O<sub>2</sub> Levels in Cellular Biology |
| title_sort | from tissue physoxia to cancer hypoxia cost effective methods to study tissue specific o sub 2 sub levels in cellular biology |
| topic | hypoxia chamber hypoxia epithelial-mesenchymal transition cancer stem cell |
| url | https://www.mdpi.com/1422-0067/23/10/5633 |
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