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Structured PDMS Chambers for Enhanced Human Neuronal Cell Activity on MEA Platforms

Journal of Bionic Engineering 2012年第1期9卷 1-10页

作者： Joose Kreutzer Laura Yla-Outinen Paula Karna Tiina Kaarela Jarno Mikkonen heli skottman Susanna Narkilahti Pasi KaUio Department of Automation Science and Engineering Tampere University of Technology Korkeakoulunkatu 3 33720 Tampere Finland BioMediTech Biokatu 10 33520 Tampere Finland NeuroGroup Institute of Biomedical Technology University of Tampere Biokatu 12 33520 Tampere Finland The Science Center of Pirkanmaa Hospital District Biokatu 12 33521 Tampere Finland Department of Psychology University of Jyvdskyld Ylistonmgientie 33 40500 Jyvdskyla Finland

Structured poly（dimethylsiloxane）（PDMS） chambers were designed and fabricated to enhance the signaling of human Embryonic Stem Cell （hESC） - derived neuronal networks on Microelectrode Array （MEA） platforms. The structured PDMS chambers enable cell seeding on restricted areas and thus, reduce the amount of needed coating materials and cells. In addition, the neuronal cells formed spontaneously active networks faster in the structured PDMS chambers than that in control chambers. In the PDMS chambers, the neuronal networks were more active and able to develop their signaling into organized signal trains faster than control cultures. The PDMS chamber design enables much more repeatable analysis and rapid growth of functional neuronal network in vitro. Moreover, due to its easy and cheap fabrication process, new configurations can be easily fabricated based on investigator requirements.

关键词： cell culturing electrical activity human embryonic stem cells microelectrode array poly(dimethylsiloxane)

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Low level of activin A secreted by fibroblast feeder cells accelerates early stage differentiation of retinal pigment epithelial cells from human pluripotent stem cells

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Stem Cell Discovery 2012年第4期2卷 176-186页

作者： Heidi Hongisto Alexandra Mikhailova Hanna Hiidenmaa Tanja Ilmarinen heli skottman Institute of Biomedical Technology University of Tampere Tampere Finland

Human pluripotent stem cells (hPSC) differentiated to retinal pigment epithelial cells (RPE) provide a promising tool for cell replacement therapies of retinal degenerative diseases. The in vitro differentiation of hPSC-RPE is still poorly understood and current differentiation protocols rely on spontaneous differentiation on fibroblast feeder cells or as floating cell aggregates in suspension. The fibroblast feeder cells may have an inductive effect on the hPSC-RPE differentiation, providing variable signals mimicking the extraocular mesenchyme that directs the differentiation in vivo. The effect of the commonly used fibroblast feeder cells on the hPSCRPE differentiation was studied by comparing suspension differentiation in standard RPEbasic (no bFGF) medium to RPEbasic medium conditioned with mouse embryonic (mEF-CM) and human foreskin (hFF-CM) fibroblast feeder cells. The fibroblast secreted factors were found to enhance early hPSC-RPE differentiation. The onset of pigmentation was faster in the conditioned media (CM) compared to RPEbasic for both human embryonic (hESC) and induced pluripotent (iPSC) stem cells, with the first pigments appearing around two weeks of differentiation. After four weeks of differentiation, CM conditions consistently contained higher number of pigmented cell aggregates. The ratio of PAX6 and MITF positive cells was quantified to be clearly higher in the CM conditions, with mEFCM containing most positive cells. The mEF cells were found to secrete low levels of activin A growth factor that is known to regulate eye field differentiation. As RPEbasic was supplemented with corresponding, low level (10 ng/ml) of recombinant human activin A, a clear increase in the hPSC-RPE differentiation was achieved. Thus, inductive effect provided by feeder cells was at least partially driven by activin A and could be substituted with a low level of recombinant growth factor in contrasts to previously reported much higher concentrations.

关键词： Retinal Pigment Epithelial Cell Human Pluripotent Stem Cell Conditioned Medium Human Foreskin Fibroblast Mouse Embryonic Fibroblast Activin A Cell Differentiation

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