Integration of 3‐D cell cultures in fluidic microsystems for biological screenings

Witte, Hartmut; Stubenrauch, Mike; Fröber, Ulrike; Fischer, R.; Voges, Danja; Hoffmann, Martin

doi:10.1002/elsc.201000045

Cited by 8 publications

(6 citation statements)

References 8 publications

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“…Our system maintained the temperature at 37 °C for more than 4 days. The measured temperature variation, ±0.3 °C, was similar to that in other studies; for instance, variations of ±0.2 °C, 27,31,32,52,53 ±0.25 °C, 18 ±0.26 °C, 10 ±0.3 °C, 54 ±0.4 °C, 55 ±0.5 °C, 56,57 and ±0.8 °C 58 have been reported. Because the intrinsic properties of fluids and cells are temperature-dependent, it is highly beneficial to prevent large temperature variations in the cell studies.…”

Section: Discussionsupporting

confidence: 89%

A Portable Microscale Cell Culture System with Indirect Temperature Control

et al. 2018

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A physiologically relevant environment is essential for successful long-term cell culturing in vitro. Precise control of temperature, one of the most crucial environmental parameters in cell cultures, increases the fidelity and repeatability of the experiments. Unfortunately, direct temperature measurement can interfere with the cultures or prevent imaging of the cells. Furthermore, the assessment of dynamic temperature variations in the cell culture area is challenging with the methods traditionally used for measuring temperature in cell culture systems. To overcome these challenges, we integrated a microscale cell culture environment together with live-cell imaging and a precise local temperature control that is based on an indirect measurement. The control method uses a remote temperature measurement and a mathematical model for estimating temperature at the desired area. The system maintained the temperature at 37±0.3 °C for more than 4 days. We also showed that the system precisely controls the culture temperature during temperature transients and compensates for the disturbance when changing the cell cultivation medium, and presented the portability of the heating system. Finally, we demonstrated a successful long-term culturing of human induced stem cell-derived beating cardiomyocytes, and analyzed their beating rates at different temperatures.

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Section: Discussionsupporting

confidence: 89%

A Portable Microscale Cell Culture System with Indirect Temperature Control

et al. 2018

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“…Numerous works showed, that MSC cultivation in the gas mixtures containing oxygen of concentrations similar to the physiological ones, is beneficial for the activation of growth and reducing the oxidative stress. Another perspective approach is the development of methods to avoid the procedure of detaching cells from a substrate, for instance, cultivation in 3D constructs or on various carriers [10], that is often technically complicated.…”

Section: Molecular and Cell Biotechnologiesmentioning

confidence: 99%

Proliferation of Wharton jelly mesenchymal stem cells, derived by preserving the cells with reduced attachment rate, under various gas conditions

Shuvalova

Кордюм

2015

Biopolym. Cell

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To estimate the proliferation rates in cultures of the human Wharton jelly mesenchymal stem cells (WJ-MSCs), obtained by the method of preservation of spontaneously detached cells, in various gas mixtures, containing physiological oxygen concentrations. Methods. Starting from the first passage, WJ-MSC were cultured for 4 subsequent passages,7 days between replating ("main" line"). At 0, 1 and 2 passage, day 3 of cultivation, conditioned media was collected and transferred to another flack with complete growth media. The attached cells from conditioned media were cultivated until the clones size and confluence (70-80 %) became sufficient for replating, and after that were subsequently passed with trypsin-EDTA solution ("side" lines). Besides the cultivation in standard condition of CO 2-incubator, the cultivation was conducted in the nitrogen-based gas mixture (3 % oxygen, 4 % carbon dioxide, 93 % nitrogen) and argonbased gas mixture (3 % oxygen, 4 % carbon dioxide, 93 % argon). At each passage, the number of cells was counted. Results. The proliferation level in "side" lines, obtained from 0 and 1 passages, had a lot of similarities with that of "main" line. We observed the trend of multiplication rate reduction in "side" lines during in vitro maintenance, similar to that in "main" line. For cultures, obtained at the passage 2, the level of proliferation was significantly lower. The cultivation in both gas mixtures with 3% O 2 concentration had beneficial effect on the level of cell multiplication: the number of cells was significantly higher. The effect of argon-based mixture was more pronounced. Conclusion. The physiologic oxygen tension allows optimizing the cultivation of WJ-MSC, obtained by the suggested method of preserving the cells with reduced attachment ability.

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“…Passage by passage the cells change their properties, lower the degree of multipotency and homing ability [9]. Therefore, various variants of cultivating MSC without any obligatory detachment are suggested, for instance, the application of culture flasks of a spe-Ó Institute of Molecular Biology and Genetics, NAS of Ukraine, 2013 cial form, which can stretch and increase the surface area with the enlargement of cell mass [10,11], or the cultivation in three-dimensional conditions on different carriers [12]. These methods are efficient, however they require additional technical possibilities, complicating the work and rather expensive.…”

mentioning

confidence: 99%

Maintenance of mesenchymal stem cells culture due to the cells with reduced attachment rate

et al. 2013

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Aim. The classic detachment techniques lead to changes in cells properties. We offer a simple method of cultivating the population of cells that avoided an influence on the surface structures. Methods. Mesenchymal stem cells (MSC) from human umbilical cord matrix were obtained and cultivated in standard conditions. While substituting the culture media by a fresh portion, the conditioned culture medium, where the cells were maintained for three days, was transferred to other culture flacks with addition of serum and growth factors. Results. In the flacks, one day after medium transfer, we observed attached cells with typical MSC morphology. The cultures originated from these cells had the same rate of surface markers expression and clonogenic potential as those replated by standard methods. Conclusions. MSC culture, derived by preserving the cells with reduced attachment ability, actually has the properties of «parent» passage. Using this method with accepted techniques of cells reseeding would allow maintaining the cells that avoided an impact on the cell surface proteins

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Integration of 3‐D cell cultures in fluidic microsystems for biological screenings

Cited by 8 publications

References 8 publications

A Portable Microscale Cell Culture System with Indirect Temperature Control

A Portable Microscale Cell Culture System with Indirect Temperature Control

Proliferation of Wharton jelly mesenchymal stem cells, derived by preserving the cells with reduced attachment rate, under various gas conditions

Maintenance of mesenchymal stem cells culture due to the cells with reduced attachment rate

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