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Electrical, chemical, and topological addressing of mammalian cells with microfabricated systems

JOURNAL OF BIOMECHANICAL ENGINEERING-TRANSACTIONS OF THE ASME 1999年第1期121卷 65-72页

作者： Kapur, R Calvert, JM Rudolph, AS George Mason Univ Ctr Bioresource Dev Fairfax VA 22030 USA USN Res Lab Ctr Biomol Sci & Engn Washington DC 20375 USA

This communication describes our work in electrical, topological, and chemical micromodification of surfaces to modulate cellular form and function. We have addressed the surface physico-chemico-mechano properties of cell culture substrates that play a role in modulating cellular behavior. Single factorial model systems have been built using techniques adapted from microlithography. The tools and techniques of microfabrication, if harnessed and used correctly, can be enabling in elucidating the underlying principles and fundamental forces driving the cell-substrate interface. Additionally, the long-term practical applications of microfabrication in medicine and biomaterial/tissue engineering lie in enabling "communication" with living cells/tissues at the cellular and subcellular levels.

关键词：生物相容性材料细胞黏附细胞培养技术/仪器和设备细胞培养技术/方法细胞骨架模型生物学单核细胞/细胞学聚合物聚硅酮类应力物理表面特性动物人类小鼠

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Adaptive control at low glucose concentration of HEK-293 cell serum-free cultures

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BIOTECHNOLOGY PROGRESS 1999年第4期15卷 608-616页

作者： Siegwart, P Côté, J Male, K Luong, JHT Perrier, M Kamen, A Natl Res Council Canada Biotechnol Res Inst Montreal PQ H4P 2R2 Canada Univ Montreal Ecole Polytech Dept Genie Chim Montreal PQ H3C 3A7 Canada

Fed-batch cultures were implemented to study the metabolism of HEK-293 cells. Glucose, measured every 30 min by a FIA biosensor system, was maintained at 1 mM throughout the culture using an adaptive nonlinear controller based on minimal process modeling. The controller performed satisfactorily at both low and high cell concentrations without the need for retuning between different culture phases. Overall, lactate production was significantly reduced by maintaining a low glucose concentration, thus decreasing the rate of glycolysis. The rates of glucose and glutamine uptake as well as the lactate and ammonia production were compared to those obtained in batch mode with an initial glucose concentration of 21 mM. Basically, three phases were observed in both culture modes. The metabolic shift from the first to the second phase was characterized by a significant reduction in glucose consumption and lactate production while maximum growth rate was maintained. The specific respiration rate appeared unchanged during the first two phases, suggesting that no change occurred in the oxidative pathway capacity. In the third phase, cell growth became slower very likely due to glutamine limitation.

关键词：氨/代谢生物反应器细胞计数细胞培养技术/仪器和设备细胞培养技术/方法细胞系培养基无血清葡萄糖/代谢葡萄糖/药理学谷氨酰胺/代谢糖酵解肾/细胞学肾/代谢动力学乳酸/生物合成人类

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Molding of deep polydimethylsiloxane microstructures for microfluidics and biological applications

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JOURNAL OF BIOMECHANICAL ENGINEERING-TRANSACTIONS OF THE ASME 1999年第1期121卷 28-34页

作者： Folch, A Ayon, A Hurtado, O Schmidt, MA Toner, M Harvard Univ Massachusetts Gen Hosp Shriners Burns HospSch Med Ctr Engn Med Boston MA 02139 USA Harvard Univ Massachusetts Gen Hosp Shriners Burns HospSch Med Surg Serv Boston MA 02139 USA MIT Microsyst Technol Labs Cambridge MA 02139 USA

Here we demonstrate the microfabrication of deep (>25 mu m) polymeric microstructures created by replica-molding polydimethylsiloxane (PDMS)from microfabricated Si substrates. The use of PDMS structures in microfluidics and biological applications is discussed. We investigated the feasibility of two methods for the microfabrication of the Si molds: deep plasma etch of silicon-on-insulator (SOI) wafers and photolithographic patterning of a spin-coated photoplastic layer. Although the SOI wafers can be patterned at higher resolution we Sound that the inexpensive photoplastic yields similar replication fidelity. The latter is mostly limited by the mechanical stability of the replicated PDMS structures. As an example, we demonstrate the selective delivery of different cell suspensions to specific locations of a tissue culture substrate resulting in micropatterns of attached cells.

关键词：生物相容性材料生物传感技术细胞黏附细胞培养技术/仪器和设备二甲基聚硅氧烷类设备设计成纤维细胞/生理学材料试验流变学聚硅酮类表面特性

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Surfaces designed to control the projected area and shape of individual cells

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JOURNAL OF BIOMECHANICAL ENGINEERING-TRANSACTIONS OF THE ASME 1999年第1期121卷 40-48页

作者： Thomas, CH Lhoest, JB Castner, DG McFarland, CD Healy, KE Northwestern Univ Sch Med Div Biol Mat Chicago IL 60611 USA Northwestern Univ Robert R McCormick Sch Engn & Appl Sci Dept Biomed Engn Evanston IL 60201 USA Univ Washington Dept Chem Engn Seattle WA 98195 USA Univ Washington Dept Bioengn Seattle WA 98195 USA

Materials with spatially resolved surface chemistry were designed to isolate individual mammalian cells to determine the influence of projected area on specific cell functions (e.g., proliferation, cytoskeletal organization). Surfaces were fabricated using a photolithographic process resulting in islands of cell binding N-(2-aminoethyl)-3-aminopropyl-trimethoxysilane (EDS) separated by a nonadhesive interpenetrating polymer network [poly (acrylamide-co-ethylene glycol);P (AAm-co-EG)]. The surfaces contained over 3800 adhesive islands/cm(2), allowing for isolation of single cells with projected areas ranging from 100 mu m(2) to 10,000 mu m(2). These surfaces provide a useful tool for researching how cell morphology and mechanical forces affect cell function.

关键词：肌动蛋白类/分析吸附生物相容性材料血蛋白质类/药代动力学细胞黏附细胞培养技术/仪器和设备细胞培养技术/方法细胞骨架设备设计水凝胶类材料试验骨关节炎髋/病理学顶骨/细胞学石英大鼠 Sprague-Dawley 硅烷类颅骨/细胞学表面特性动物人类大鼠

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Stability of oxygen deprivation in glass culture vessels facilitates fast reproducible cell death to cortical neurons under simulated ischemia

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ANALYTICAL BIOCHEMISTRY 2002年第1期306卷 149-152页

作者： Munns, SE Arthur, PG Univ Western Australia Dept Biochem Crawley WA 6009 Australia Western Australian Inst Med Res Perth WA Australia

Glass petri dishes provide tight control of oxygen concentration during continuous nitrogen gassing to cultured cells. This control has led to a quick and highly reproducible simulated ischemic model of cortical neuron death in vitro. Twenty-five minutes of simulated ischemia was required for 92% death in cortical neuron cultures on glass. The contamination of leached oxygen from plastic culture vessels, in comparative experiments, necessitates longer times of simulated ischemia (120 min) to achieve similar cell death. Steady-state release of oxygen from plastic petri dishes was observed for up to 60 min of nitrogen gassing, providing a metabolic environment contrary to that of anoxia. Further investigations with models of simulated ischemia in glass culture vessels could answer some criticisms of in vitro models and their relevance to in vivo situations (10).

关键词：脑缺血/代谢脑缺血/病理学细胞培养技术/仪器和设备细胞培养技术/方法细胞死亡/药物作用细胞低氧/生理学细胞存活/药物作用细胞培养的大脑皮质/药物作用大脑皮质/胚胎学大脑皮质/病理学玻璃糖酵解 L-乳酸脱氢酶/分析神经元/药物作用神经元/代谢神经元/病理学氧/代谢氧/药理学可重复性结果动物大鼠

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A new alpha-particle irradiator with absolute dosimetric determination

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RADIATION RESEARCH 2000年第1期153卷 9-15页

作者： Soyland, C Hassfjell, SP Steen, HB Norwegian Radium Hosp Dept Biophys N-0310 Oslo Norway Univ Oslo Dept Chem N-0315 Oslo Norway

A new experimental setup for uniform alpha-particle irradiation of cells in vitro is described. The alpha-particle irradiator is based on a radioactive Pb-212/Bi-212 source. In the experimental setup proposed, cells are grown directly on a polylysine-coated track-etch material that forms the base of custom-made cell dishes. Alpha-particle irradiation is done through the base of the dish, Immediately prior to irradiation, the cell dish is scanned under a microscope, and images of cells with the corresponding coordinates are saved, After irradiation and after the biological end point under study has been determined, the cell dish is etched to develop alpha-particle tracks in the dish base. A microscope image series of alpha-particle track images is obtained by accurately revisiting every original (preirradiation) cell position in the track-etched dish, The number of alpha-particle traversals of each individual cell is scored by mapping images of alpha-particle tracks onto the images of cells recorded prior to irradiation. The uncertainty of the alpha-particle hit determination is 0.9 mu m. The procedure described thus presents a method for radiobiological experiments with absolute, rather than statistical, cell dosimetry, (C) 2000 by Radiation Research Society.

关键词：空气 α粒子 β射线铋细胞培养技术/仪器和设备细胞/辐射效应细胞培养的/辐射效应设备设计铅放射性同位素传能线密度肺/细胞学肺/辐射效应聚对苯二甲酸乙酯类/辐射效应辐射防护/方法放射测量术/仪器和设备放射测量术/方法光谱分析钍人类

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Animal-cell damage in sparged bioreactors

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TRENDS IN BIOTECHNOLOGY 2000年第10期18卷 420-432页

作者： Chisti, Y Univ Almeria Dept Chem Engn E-04071 Almeria Spain

The gas sparging of culture broth causes damage to suspended animal cells. However, despite this, sparged bioreactors remain the preferred means of cell culture because sparging is a robust method of supplying oxygen, especially on a large scale. This article examines the underlying mechanisms involved in bubble-associated cell damage and the methods available for controlling such damage.

关键词：生物反应器血液/代谢细胞培养技术/仪器和设备细胞培养技术/方法培养基动力学模型理论氧/代谢泊洛沙姆/药理学聚乙烯二醇类/代谢表面活性剂/药理学动物

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