Design and Characterization of a Sensorized Microfluidic  Cell-Culture System with Electro-Thermal Micro-Pumps and Sensors for Cell Adhesion, Oxygen, and pH on a Glass Chip

Bonk, Sebastian M.; Stubbe, Marco; Buehler, Sebastian M.; Tautorat, Carsten; Baumann, Werner; Klinkenberg, E.-D.; Gimsa, Jan

doi:10.3390/bios5030513

Cited by 31 publications

(33 citation statements)

References 57 publications

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“…Although current research mainly aims at establishing a dynamic culture to increase oxygen supply [14] , we assume that this problem may be less important than the removal of CO 2 to avoid acidification by carbonate. With MC3T3 osteoblasts, we showed that a feeding medium-pH of 7.8 helped uphold the pH above 7.0 in confined microfluidic cell culture systems for longer medium exchange periods [15] . We speculate that elevated pH may provide a simple way to avoid adverse effects on cell proliferation and cell vitality during static cell culturing in 3D scaffolds, not only for osteoblasts.…”

Section: Introductionmentioning

confidence: 92%

Increased osteoblast viability at alkaline pH in vitro provides a new perspective on bone regeneration

Galow

Rebl

Koczan

et al. 2017

Biochemistry and Biophysics Reports

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102

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We investigated the effects of alkaline pH on developing osteoblasts. Cells of the osteoblast-like cell line MC3T3-E1 were initially cultured for six days in HEPES-buffered media with pH ranging from 7.2 to 9.0. Cell count, cellular WST-1 metabolism, and ATP content were analyzed. The three parameters showed a pH optimum around pH 8.4, exceeding the recommended buffer range of HEPES at the alkaline flank. Therefore, only pH 7.2, 7.4, 7.8, and 8.4 media were used in more elaborate, daily investigations to reduce the effects of pH change within the pH control intervals of 24 h. All parameters exhibited similar pH behaviors, roughly showing increases to 130% and 230% at pH 7.8 and 8.4, as well as decreases to 70% at pH 7.2 when using the pH 7.4 data for reference. To characterize cell differentiation and osteoblastic cell function, cells were cultured at pH 7.4 and under alkaline conditions at pH 7.8 and 8.4 for 14 days. Gene expression and mineralization were evaluated using microarray technology and Alizarin staining. Under alkaline conditions, ATF4, a regulator for terminal differentiation and function as well as DMP1, a potential marker for the transition of osteoblasts into osteocytes, were significantly upregulated, hinting at an accelerated differentiation process. After 21 days, significant mineralization was only detected at alkaline pH. We conclude that elevated pH is beneficial for the cultivation of bone cells and may also provide therapeutic value in bone regeneration therapies.

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Section: Introductionmentioning

confidence: 92%

Increased osteoblast viability at alkaline pH in vitro provides a new perspective on bone regeneration

Galow

Rebl

Koczan

et al. 2017

Biochemistry and Biophysics Reports

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102

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“…The glass metabolic chip ( Figure 2 B) was a further development of the chip created by Bonk et al [ 51 ]. The microfluidic structure was imprinted in a poly-dimethyl-siloxane (PDMS) layer on the bottom side of a poly-methyl-methacrylate (Plexiglas ® GS, Acrylics Ltd. Niederfischbach, Germany) lid containing four microfluidic ports with tube connectors.…”

Section: Sensorized Glass Chipsmentioning

confidence: 99%

“…After these tests were completed, cell culturing was conducted with the lower left and top right connectors being used as the inlet and outlet, respectively. The inlet and outlet were connected with Teflon (PTFE) tubes to the medium flask and an external peristaltic pump [ 51 ]. The other two connectors were sealed.…”

Section: Sensorized Glass Chipsmentioning

confidence: 99%

Cell Monitoring and Manipulation Systems (CMMSs) based on Glass Cell-Culture Chips (GC3s)

et al. 2016

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We developed different types of glass cell-culture chips (GC3s) for culturing cells for microscopic observation in open media-containing troughs or in microfluidic structures. Platinum sensor and manipulation structures were used to monitor physiological parameters and to allocate and permeabilize cells. Electro-thermal micro pumps distributed chemical compounds in the microfluidic systems. The integrated temperature sensors showed a linear, Pt1000-like behavior. Cell adhesion and proliferation were monitored using interdigitated electrode structures (IDESs). The cell-doubling times of primary murine embryonic neuronal cells (PNCs) were determined based on the IDES capacitance-peak shifts. The electrical activity of PNC networks was detected using multi-electrode arrays (MEAs). During seeding, the cells were dielectrophoretically allocated to individual MEAs to improve network structures. MEA pads with diameters of 15, 20, 25, and 35 µm were tested. After 3 weeks, the magnitudes of the determined action potentials were highest for pads of 25 µm in diameter and did not differ when the inter-pad distances were 100 or 170 µm. Using 25-µm diameter circular oxygen electrodes, the signal currents in the cell-culture media were found to range from approximately −0.08 nA (0% O2) to −2.35 nA (21% O2). It was observed that 60-nm thick silicon nitride-sensor layers were stable potentiometric pH sensors under cell-culture conditions for periods of days. Their sensitivity between pH 5 and 9 was as high as 45 mV per pH step. We concluded that sensorized GC3s are potential animal replacement systems for purposes such as toxicity pre-screening. For example, the effect of mefloquine, a medication used to treat malaria, on the electrical activity of neuronal cells was determined in this study using a GC3 system.

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“…The analysis of real samples implies the determination of several ions. Analytes of interest are related to acidification, respiration and some ions, such as sodium and potassium among the most general ones [16,17]. Oxygen consumption [18] and acidification [19] are both indicative of cell metabolism.…”

Section: Introductionmentioning

confidence: 99%

Miniaturized multiparametric flexible platform for the simultaneous monitoring of ionic: Application in real urine

Moya

Illa

Gíménez

et al. 2018

Sensors and Actuators B: Chemical

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International audienceBiomonitoring is a research topic that has largely relied on cell culture systems. Recently, the development of “Organ-on-a-Chip” (OC) platforms and the need for a continuous monitoring of these systems has increased its interest. However, the biomonitorization in these systems is still at its infancy due to the difficulty to adapt the sensors to microfluidic OC systems. In this work we have fabricated a modular, versatile, scalable, multi-analyte sensing platform, which integrates dissolved oxygen (DO), Na+, K+ and pH sensors for monitoring their concentrations in small volume of biological fluids. The platform is fabricated by means of rapid prototyping techniques using polymeric substrates, and incorporates all the necessary elements to measure the four analytes. It was designed with four gold sensing electrodes, and integrates a gold counter electrode and an Ag/AgCl pseudo-reference electrode for the electrochemical measurements in small volumes (0.83 μL) of samples. The sensors featured good sensitivities of 3.60 ± 0.2 nA (mg L−1) for DO and 69 ± 1 mV decade−1 for pH. Na+ and K+ μISE exhibited sensitivities of 57 ± 1 mV decade−1 and 52 ± 2 mV decade−1, and low limits of detection 5 × 10−6 M and 0.5 × 10−5 M respectively. This platform allows the dynamic measurement of the biological fluids parameters simultaneously, in real time and with a rapid response. The versatility of the platform allows its adaptation in any microfluidic cells culture systems. The novel multi-sensor platform has been validated in controlled buffers and with artificial urine (AU). A proof-of-concept using real mice urine (RU) has been carried out, demonstrating the good behaviour of the multi-sensing platform

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Design and Characterization of a Sensorized Microfluidic Cell-Culture System with Electro-Thermal Micro-Pumps and Sensors for Cell Adhesion, Oxygen, and pH on a Glass Chip

Cited by 31 publications

References 57 publications

Increased osteoblast viability at alkaline pH in vitro provides a new perspective on bone regeneration

Increased osteoblast viability at alkaline pH in vitro provides a new perspective on bone regeneration

Cell Monitoring and Manipulation Systems (CMMSs) based on Glass Cell-Culture Chips (GC3s)

Miniaturized multiparametric flexible platform for the simultaneous monitoring of ionic: Application in real urine

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