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"lastName": "January" }, { "creatorType": "author", "firstName": "David J.", "lastName": "Beebe" } ], "abstractNote": "Microfluidics is emerging as a promising platform for cell culture, enabling increased microenvironment control and potential for integrated analysis compared to conventional macroculture systems such as well plates and petri dishes. To advance the use of microfluidic devices for cell culture, it is necessary to better understand how miniaturization affects cell behavior. In particular, microfluidic devices have significantly higher surface-area-to-volume ratios than conventional platforms, resulting in lower volumes of media per cell, which can lead to cell stress. We investigated cell stress under a variety of culture conditions using three cell lines: parental HEK (Human Embryonic Kidney) cells and transfected HEK cells that stably express wild type (WT) and mutant (G601S) human ether-a-go-go related gene (hERG) potassium channel protein. These three cell lines provide a unique model system through which to study cell-type-specific responses in microculture because mutant hERG is known to be sensitive to environmental conditions, making its expression a particularly sensitive readout through which to compare macro- and microculture. While expression of WT-hERG was similar in microchannel and well culture, the expression of mutant G601S-hERG was reduced in microchannels. Expression of the endoplasmic reticulum (ER) stress marker Immunoglobulin Binding Protein (BiP) was upregulated in all three cell lines in microculture. Using BiP expression, glucose consumption, and lactate accumulation as readouts we developed methods for reducing ER stress including properly increasing the frequency of media replacement, reducing cell seeding density, and adjusting the serum concentration and buffering capacity of culture medium. Indeed, increasing the buffering capacity of culture medium or frequency of media replacement partially restored the expression of the G601S-hERG in microculture. This work illuminates how biochemical properties of cells differ in macro- and microculture and suggests strategies that can be used to modify cell culture protocols for future studies involving miniaturized culture platforms.", "publicationTitle": "Analytical chemistry", "publisher": "", "place": "", "date": "2013-2-5", "volume": "85", "issue": "3", "section": "", "partNumber": "", "partTitle": "", "pages": "1562-1570", "series": "", "seriesTitle": "", "seriesText": "", "journalAbbreviation": "Anal Chem", "DOI": "10.1021/ac3027228", "citationKey": "", "url": "http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3565071/", "accessDate": "2016-01-03T20:49:34Z", "PMID": "", "PMCID": "", "ISSN": "0003-2700", "archive": "", "archiveLocation": "", "shortTitle": "Effect of microculture on cell metabolism and biochemistry", "language": "", "libraryCatalog": "PubMed Central", "callNumber": "", "rights": "", "extra": "PMID: 23327437\nPMCID: PMC3565071", "tags": [], "collections": [], "relations": {}, 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"abstractNote": "It is now well documented that apoptosis represents the prevalent mode of cell death in hybridoma cultures. Apoptotic or programmed cell death occurs spontaneously in late exponential phase of batch cultures. Until lately, no specific triggering factors had been identified. Recently, we observed that glutamine, cystine or glucose deprivation induced apoptosis in both hybridoma and myeloma cell lines whereas accumulation of toxic metabolites induced necrotic cell death in these cells. Other triggering factors such as oxygen deprivation might also be responsible for induction of apoptosis. In the present study, induction of cell death by exposure to anoxia was examined in batch culture of the SP2/0-derived hybridoma D5 clone. The mode of cell death was studied by morphological examination of acridine orange-ethidium bromide stained cells in a 1.5 L bioreactor culture grown under anoxic conditions for 75 hours. Under such conditions, viable cell density levelled off rapidly and remained constant for 25 hours. After 45 hours of anoxia, cell viability had decreased to 30% and the dead cell population was found to be 90% apoptotic. In terms of cellular metabolism, anoxia resulted in an increase in the utilization rates of glucose and arginine, and in a decrease in the utilization rate of glutamine. The lactate production rate and the yield of lactate on glucose increased significantly while the MAb production rate decreased. These results demonstrate that glycolysis becomes the main source of energy under anoxic conditions. Cells incubated for 10 hours or less under anoxic conditions were able to recuperate almost immediately and displayed normal growth rates when reincubated in oxic conditions whereas cells incubated for 22 hours or more displayed reduced growth rates. Nonetheless, even after 22 h or 29 h of anoxia, cells reincubated in oxic conditions showed no further progression into apoptosis. Therefore, upon removal of the triggering signal, induction of apoptosis ceased.", "publicationTitle": "Cytotechnology", "publisher": "", "place": "", "date": "1994-02", "volume": "15", "issue": "1-3", "section": "", "partNumber": "", "partTitle": "", "pages": "117-128", "series": "", "seriesTitle": "", "seriesText": "", "journalAbbreviation": "Cytotechnology", "DOI": "10.1007/BF00762386", "citationKey": "", "url": "http://link.springer.com/article/10.1007/BF00762386", "accessDate": "2016-01-03T01:52:29Z", "PMID": "", "PMCID": "", "ISSN": "0920-9069, 1573-0778", "archive": "", "archiveLocation": "", "shortTitle": "", "language": "en", "libraryCatalog": "link.springer.com", "callNumber": "", "rights": "", "extra": "", "tags": [ { "tag": "Animal cell culture", "type": 1 }, { "tag": "Biochemistry, general", "type": 1 }, { "tag": "Biomedicine general", "type": 1 }, { "tag": "Cell Biology", "type": 1 }, { "tag": "Proteomics", "type": 1 }, { "tag": "anoxia", "type": 1 }, { "tag": 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