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1. 发明授权

US09163216B1 Method for culturing skeletal muscle for tissue engineering 有权
标题翻译：组织工程骨骼肌培养方法
公开(公告)号：US09163216B1
公开(公告)日：2015-10-20
申请号：US12765399
申请日：2010-04-22
申请人： James J. Hickman , Mainak Das , John W. Rumsey
发明人： James J. Hickman , Mainak Das , John W. Rumsey
IPC分类号： C12N5/077 , C12N5/0793
CPC分类号： C12N5/0652 , C12N5/0619 , C12N5/0658 , C12N2500/05 , C12N2500/46 , C12N2500/90 , C12N2500/99 , C12N2501/40 , C12N2502/1335 , C12N2533/30
摘要： The invention provides a nutrient medium composition and associated methods for lengthening the useful life of a culture of muscle cells. Disclosed is a method of culturing mammalian muscle cells, including preparing one or more carriers coated with a covalently bonded monolayer of trimethoxy-silylpropyl-diethylenetriamine (DETA); verifying DETA monolayer formation by one or more associated optical parameters; suspending isolated fetal rat skeletal muscle cells in serum-free medium according to medium composition 1; plating the suspended cells onto the prepared carriers at a predetermined density; leaving the carriers undisturbed for cells to adhere to the DETA monolayer; covering the carriers with a mixture of medium 1 and medium 2; and incubating. A cell nutrient medium composition includes Neurobasal, an antibiotic-antimycotic composition, cholesterol, human TNF-alpha, PDGF BB, vasoactive intestinal peptides, insulin-like growth factor 1, NAP, r-Apolipoprotein E2, purified mouse Laminin, beta amyloid, human tenascin-C protein, rr-Sonic hedgehog Shh N-terminal, and rr-Agrin C terminal.
摘要翻译：本发明提供了一种营养培养基组合物和相关方法，用于延长肌肉细胞培养物的使用寿命。公开了一种培养哺乳动物肌肉细胞的方法，包括制备一种或多种涂覆有共价键合的三甲氧基 - 甲硅烷基丙基 - 二亚乙基三胺（DETA）单层的载体; 通过一个或多个相关联的光学参数验证DETA单层形成; 根据培养基组成1，在无血清培养基中悬浮分离的胎鼠大鼠骨骼肌细胞; 以预定的密度将悬浮细胞电镀到制备的载体上; 使载体不受干扰以使细胞粘附到DETA单层上; 用介质1和介质2的混合物覆盖载体; 并孵化。细胞营养培养基组合物包括Neurobasal，抗生素 - 抗真菌组合物，胆固醇，人TNF-α，PDGF BB，血管活性肠肽，胰岛素样生长因子1，NAP，r-载脂蛋白E2，纯化的小鼠层粘连蛋白，β淀粉样蛋白，人腱生蛋白-C蛋白，rr-Sonic刺猬Shh N末端和rr-Agrin C端。

2. 发明授权

US08815584B1 Method of co-culturing mammalian muscle cells and motoneurons 有权
标题翻译：共培养哺乳动物肌肉细胞和运动神经元的方法
公开(公告)号：US08815584B1
公开(公告)日：2014-08-26
申请号：US12765996
申请日：2010-04-23
申请人： James J. Hickman , Mainak Das
发明人： James J. Hickman , Mainak Das
IPC分类号： C12N5/07 , C12N11/00 , C12N11/16
CPC分类号： C12N5/0619 , C12N5/0658 , C12N2500/90 , C12N2502/088 , C12N2502/1335
摘要： The invention provides a method of co-culturing mammalian muscle cells and mammalian motoneurons. The method comprises preparing one or more carriers coated with a covalently bonded monolayer of trimethoxysilylpropyl diethylenetriamine (DETA); suspending isolated fetal mammalian skeletal muscle cells in serum-free medium according to medium composition 1; suspending isolated fetal mammalian spinal motoneurons in serum-free medium according to medium composition 1; plating the suspended muscle cells onto the one or more carriers at a predetermined density and allowing the muscle cells to attach; plating the suspended motoneurons at a predetermined density onto the one or more carriers and allowing the motoneurons to attach; covering the one or more carriers with a mixture of medium composition 1 and medium composition 2; and incubating the carriers covered in the media mixture.
摘要翻译：本发明提供了共培养哺乳动物肌肉细胞和哺乳动物运动神经元的方法。该方法包括制备涂覆有共价键合的三甲氧基甲硅烷基丙基二亚乙基三胺（DETA）单层的一种或多种载体; 根据培养基组成1将分离的胎儿哺乳动物骨骼肌细胞悬浮于无血清培养基中; 根据培养基组成1，在无血清培养基中悬浮胎儿哺乳动物脊髓运动神经元; 以预定的密度将悬浮的肌肉细胞电镀到一个或多个载体上并允许肌肉细胞附着; 将预定密度的悬浮运动神经元电镀到一个或多个载体上并允许运动神经元附着; 用培养基组合物1和培养基组合物2的混合物覆盖一种或多种载体; 并培养介质混合物中覆盖的载体。

3. 发明申请

US20120128639A1 In Vitro Production Of Oligodendrocytes From Human Umbilical Cord Stem Cells 审中-公开
标题翻译：从人脐带干细胞体外产生少突胶质细胞
公开(公告)号：US20120128639A1
公开(公告)日：2012-05-24
申请号：US13322903
申请日：2010-05-28
申请人： James J. Hickman , Hedvika Davis
发明人： James J. Hickman , Hedvika Davis
IPC分类号： A61K35/30 , A61P25/00 , C12N5/079
CPC分类号： C12N5/0622 , C12N2500/90 , C12N2501/01 , C12N2501/11 , C12N2501/115 , C12N2501/135 , C12N2501/70 , C12N2501/81 , C12N2501/91 , C12N2506/025 , C12N2506/1369 , C12N2506/1392
摘要： The invention provides a method of producing oligodendrocytes by in vitro differentiation of human multi-potent progenitor cells (MLPCs). The method comprises culturing isolated MLPCs on a first surface in a serum-free defined culture medium; replacing the culture medium with serum-free culture medium supplemented with bFGF, EGF and PDGF-AA for approximately 24 hours; changing the cultured MLPCs into the supplemented serum-free culture medium further supplemented with differentiation factors norepinephrine, forskolin. and K252a; establishing a 3D environment by covering the culture with a second surface opposite and spaced apart from the first surface, so as to contain the MLPCs therebetween; and continuing to culture until a majority of the MLPCs have differentiated into oligodendrocytes. Additionally included is a method of treatment for a subject afflicted by a disease characterized by central or peripheral nervous system demyelination, the method comprising transplanting into the subject oligodendrocytes produced according to the method disclosed.
摘要翻译：本发明提供了通过体外分化人多能祖细胞（MLPC）产生少突胶质细胞的方法。该方法包括在无血清定义培养基的第一表面培养分离的MLPC; 用补充有bFGF，EGF和PDGF-AA的无血清培养基代替培养基约24小时; 将培养的MLPC变成补充的无血清培养基，进一步补充分化因子去甲肾上腺素，毛喉素。和K252a; 通过用与第一表面相对且间隔开的第二表面覆盖培养来建立3D环境，以便在其间容纳MLPC; 并继续培养，直到大多数MLPC已经分化为少突胶质细胞。另外包括治疗由以中枢神经系统或周围神经系统脱髓鞘为特征的疾病所折磨的受试者的方法，该方法包括将根据所公开的方法产生的少突胶质细胞移植到受试者中。

4. 发明授权

US07266457B1 High throughput functional genomics 有权
标题翻译：高通量功能基因组学
公开(公告)号：US07266457B1
公开(公告)日：2007-09-04
申请号：US09575377
申请日：2000-05-22
申请人： James J. Hickman
发明人： James J. Hickman
IPC分类号： G06F19/00 , C12N15/00 , G01N33/53 , C12M1/34
CPC分类号： B82Y30/00 , C12Q1/001 , G01N33/48707 , G01N33/54373
摘要： This invention focuses on the marriage of solid-state electronics and neuronal function to create a new high-throughput electrophysiological assay to determine a compound's acute and chronic effect on cellular function. Electronics, surface chemistry, biotechnology, and fundamental neuroscience are integrated to provide an assay where the reporter element is an array of electrically active cells. This innovative technology can be applied to neurotoxicity, and to screening compounds from combinatorial chemistry, gene function analysis, and basic neuroscience applications. The system of the invention analyzes how the action potential is interrupted by drugs or toxins. Differences in the action potentials are due to individual toxins acting on different biochemical pathways, which in turn affects different ion channels, thereby changing the peak shape of the action potential differently for each toxin. Algorithms to analyze the action potential peak shape differences are used to indicate the pathway(s) affected by the presence of a new drug or compound; from that, aspects of its function in that cell are deduced. This observation can be exploited to determine the functional category of biochemical action of an unknown compound. An important aspect of the invention is surface chemistry that permits establishment of a high impedance seal between cell and a metal microelectrode. This seal recreates the interface that enables functional patch-clamp electrophysiology with glass micropipettes, and allows extracellular electrophysiology on a microelectrode array. Thus, the invention teaches the feasibility of using living cells as diagnostics for high throughput real-time assays of cell function.
摘要翻译：本发明专注于固态电子学和神经元功能的结合，以创建新的高通量电生理测定法，以确定化合物对细胞功能的急性和慢性影响。电子学，表面化学，生物技术和基础神经科学被整合，以提供报告元素是电活性细胞阵列的测定。这种创新技术可以应用于神经毒性，并从组合化学，基因功能分析和基础神经科学应用中筛选化合物。本发明的系统分析动作电位如何被药物或毒素中断。动作电位的差异是由于各种毒素作用于不同的生物化学途径，这反过来又影响不同的离子通道，从而改变每种毒素的动作电位的峰值形状。用于分析动作电位峰形差异的算法用于指示受新药或化合物存在影响的途径; 从此，推导出其在该单元中的功能的方面。可以利用这一观察来确定未知化合物的生物化学作用的功能类别。本发明的一个重要方面是允许在电池和金属微电极之间建立高阻抗密封的表面化学。该密封件重新创建了使用玻璃微量移液器实现功能性膜片钳电生理学的界面，并允许微电极阵列上的细胞外电生理学。因此，本发明教导了使用活细胞作为细胞功能的高通量实时测定的诊断的可行性。

5. 发明授权

US5342737A High aspect ratio metal microstructures and method for preparing the same 失效
标题翻译：高纵横比金属微结构及其制备方法
公开(公告)号：US5342737A
公开(公告)日：1994-08-30
申请号：US874403
申请日：1992-04-27
申请人： Jacque H. Georger, Jr. , Martin C. Peckerar , Milton L. Rebbert , Jeffrey M. Calvert , James J. Hickman
发明人： Jacque H. Georger, Jr. , Martin C. Peckerar , Milton L. Rebbert , Jeffrey M. Calvert , James J. Hickman
IPC分类号： G03F1/00 , G03F7/00 , G03F7/40 , G03C5/00
CPC分类号： G03F7/00 , G03F1/22 , G03F7/40 , Y10S414/123 , Y10S428/923 , Y10T428/12292
摘要： High aspect ratio metal microstructures may be prepared by a method involving(i) forming a layer of a photoresist on a substrate;(ii) exposing the layer to actinic radiation in an imagewise manner and developing the exposed layer to obtain a surface which contains regions having no remaining photoresist and regions covered with photoresist;(iii) metallizing the surface to form a layer of metal on the region of the surface having no remaining photoresist and on the sides of the regions of photoresist remaining on the surface; and(iv) optionally, stripping the photoresist remaining on the surface.Such microstructures are useful as electron emitters, anisotropic high dielectric interconnects, masks for x-ray photolithography, carriers for the controlled release of active agents, and ultramicroelectrode arrays.
摘要翻译：高纵横比金属微结构可以通过以下方法制备：（i）在基底上形成光致抗蚀剂层; （ii）以成像方式将层暴露于光化辐射并显影曝光层以获得包含没有剩余光致抗蚀剂区域和用光刻胶覆盖的区域的表面; （iii）使表面金属化以在表面区域上形成金属层，该区域上没有剩余的光致抗蚀剂，并且在保留在表面上的光致抗蚀剂区域的侧面上; 和（iv）任选地，剥离残留在表面上的光致抗蚀剂。这种微结构可用作电子发射体，各向异性高介电互连，用于X射线光刻的掩模，用于控制释放活性剂的载体和超微电极阵列。

6. 发明授权

US07734426B2 High throughput functional genomics 失效
标题翻译：高通量功能基因组学
公开(公告)号：US07734426B2
公开(公告)日：2010-06-08
申请号：US11840041
申请日：2007-08-16
申请人： James J. Hickman
发明人： James J. Hickman
IPC分类号： G06F19/00 , C12N15/00 , G01N33/53 , C12M1/34
CPC分类号： B82Y30/00 , C12Q1/001 , G01N33/48707 , G01N33/54373
摘要： This invention focuses on the marriage of solid-state electronics and neuronal function to create a new high-throughput electrophysiological assay to determine a compound's acute and chronic effect on cellular function. Electronics, surface chemistry, biotechnology, and fundamental neuroscience are integrated to provide an assay where the reporter element is an array of electrically active cells. This innovative technology can be applied to neurotoxicity, and to screening compounds from combinatorial chemistry, gene function analysis, and basic neuroscience applications. The system of the invention analyzes how the action potential is interrupted by drugs or toxins. Differences in the action potentials are due to individual toxins acting on different biochemical pathways, which in turn affects different ion channels, thereby changing the peak shape of the action potential differently for each toxin. Algorithms to analyze the action potential peak shape differences are used to indicate the pathway(s) affected by the presence of a new drug or compound; from that, aspects of its function in that cell are deduced. This observation can be exploited to determine the functional category of biochemical action of an unknown compound. An important aspect of the invention is surface chemistry that permits establishment of a high impedance seal between cell and a metal microelectrode. This seal recreates the interface that enables functional patch-clamp electrophysiology with glass micropipettes, and allows extracellular electrophysiology on a microelectrode array. Thus, the invention teaches the feasibility of using living cells as diagnostics for high throughput real-time assays of cell function.
摘要翻译：本发明专注于固态电子学和神经元功能的结合，以创建新的高通量电生理测定法，以确定化合物对细胞功能的急性和慢性影响。电子学，表面化学，生物技术和基础神经科学被整合，以提供报告元素是电活性细胞阵列的测定。这种创新技术可以应用于神经毒性，并从组合化学，基因功能分析和基础神经科学应用中筛选化合物。本发明的系统分析动作电位如何被药物或毒素中断。动作电位的差异是由于各种毒素作用于不同的生物化学途径，这反过来又影响不同的离子通道，从而改变每种毒素的动作电位的峰值形状。用于分析动作电位峰形差异的算法用于指示受新药或化合物存在影响的途径; 从此，推导出其在该单元中的功能的方面。可以利用这一观察来确定未知化合物的生物化学作用的功能类别。本发明的一个重要方面是允许在电池和金属微电极之间建立高阻抗密封的表面化学。该密封件重新创建了使用玻璃微量移液器实现功能性膜片钳电生理学的界面，并允许微电极阵列上的细胞外电生理学。因此，本发明教导了使用活细胞作为细胞功能的高通量实时测定的诊断的可行性。

7. 发明授权

US5223117A Two-terminal voltammetric microsensors 失效
标题翻译：两端电压显微镜
公开(公告)号：US5223117A
公开(公告)日：1993-06-29
申请号：US695254
申请日：1991-05-03
申请人： Mark S. Wrighton , James J. Hickman , Paul E. Laibinis , David Ofer , Chad A. Mirkin , James R. Valentine , George M. Whitesides
发明人： Mark S. Wrighton , James J. Hickman , Paul E. Laibinis , David Ofer , Chad A. Mirkin , James R. Valentine , George M. Whitesides
IPC分类号： G01N27/416
CPC分类号： B82Y30/00 , B82Y15/00 , G01N27/3277
摘要： Self-assembly of a chemically insensitive redox material, such as ferrocenyl thiol, and a chemically sensitive redox material, such as a quinone thiol, onto microelectrodes forms the basis for a two-terminal, voltammetric microsensor having reference and sensor functions on the same electrode. Detection is based on measuring the potential difference of current peaks for oxidation and reduction of the reference (ferrocene) and indicator (quinone) in aqueous electrolyte in a two-terminal, linear sweep voltammogram using a counterelectrode of relatively large surface area. Use of microelectrodes modified with monolayer coverages of reference and indicator molecules minimizes the size of the counterelectrode and the perturbation of the solution interrogated. Key advantages are that the sensor requires no separate reference electrode and the sensor functions as long as current peaks can be located for reference and indicator molecules.
摘要翻译：化学不敏感的氧化还原材料（例如二茂铁基硫醇）和化学敏感的氧化还原材料（例如醌硫醇）自组装到微电极上形成了在相同电极上具有参考和传感器功能的两端伏安微传感器的基础。检测是基于使用相对较大的表面积的反电极测量在两端线性扫描伏安图中的水性电解质中氧化和还原参考（二茂铁）和指示剂（醌）的电流峰的潜在差异。使用用参考和指示剂分子的单层覆盖物修饰的微电极最小化了反电极的尺寸和被询问的溶液的扰动。主要优点是传感器不需要单独的参比电极，传感器的作用只要能够为参考和指示剂分子定位电流峰值。

8. 发明申请

US20080124789A1 High Throughput Functional Genomics 失效
标题翻译：高通量功能基因组学
公开(公告)号：US20080124789A1
公开(公告)日：2008-05-29
申请号：US11840041
申请日：2007-08-16
申请人： James J. Hickman
发明人： James J. Hickman
IPC分类号： C12M1/34 , G06G7/48
CPC分类号： B82Y30/00 , C12Q1/001 , G01N33/48707 , G01N33/54373
摘要： This invention focuses on the marriage of solid-state electronics and neuronal function to create a new high-throughput electrophysiological assay to determine a compound's acute and chronic effect on cellular function. Electronics, surface chemistry, biotechnology, and fundamental neuroscience are integrated to provide an assay where the reporter element is an array of electrically active cells. This innovative technology can be applied to neurotoxicity, and to screening compounds from combinatorial chemistry, gene function analysis, and basic neuroscience applications. The system of the invention analyzes how the action potential is interrupted by drugs or toxins. Differences in the action potentials are due to individual toxins acting on different biochemical pathways, which in turn affects different ion channels, thereby changing the peak shape of the action potential differently for each toxin. Algorithms to analyze the action potential peak shape differences are used to indicate the pathway(s) affected by the presence of a new drug or compound; from that, aspects of its function in that cell are deduced. This observation can be exploited to determine the functional category of biochemical action of an unknown compound. An important aspect of the invention is surface chemistry that permits establishment of a high impedance seal between cell and a metal microelectrode. This seal recreates the interface that enables functional patch-clamp electrophysiology with glass micropipettes, and allows extracellular electrophysiology on a microelectrode array. Thus, the invention teaches the feasibility of using living cells as diagnostics for high throughput real-time assays of cell function.
摘要翻译：本发明专注于固态电子学和神经元功能的结合，以创建新的高通量电生理测定法，以确定化合物对细胞功能的急性和慢性影响。电子学，表面化学，生物技术和基础神经科学被整合，以提供报告元素是电活性细胞阵列的测定。这种创新技术可以应用于神经毒性，并从组合化学，基因功能分析和基础神经科学应用中筛选化合物。本发明的系统分析动作电位如何被药物或毒素中断。动作电位的差异是由于各种毒素作用于不同的生物化学途径，这反过来又影响不同的离子通道，从而改变每种毒素的动作电位的峰值形状。用于分析动作电位峰形差异的算法用于指示受新药或化合物存在影响的途径; 从此，推导出其在该单元中的功能的方面。可以利用这一观察来确定未知化合物的生物化学作用的功能类别。本发明的一个重要方面是允许在电池和金属微电极之间建立高阻抗密封的表面化学。该密封件重新创建了使用玻璃微量移液器实现功能性膜片钳电生理学的界面，并允许微电极阵列上的细胞外电生理学。因此，本发明教导了使用活细胞作为细胞功能的高通量实时测定的诊断的可行性。

9. 发明授权

US06654729B1 Neuroelectric computational devices and networks 有权
标题翻译：神经电学计算设备和网络
公开(公告)号：US06654729B1
公开(公告)日：2003-11-25
申请号：US09927746
申请日：2001-08-09
申请人： James J. Hickman , Joseph J. Pancrazio
发明人： James J. Hickman , Joseph J. Pancrazio
IPC分类号： G06N302
CPC分类号： B82Y30/00 , G06N3/061
摘要： A neuroelectric device is defined by a substrate having a neuronal cell provided thereon, a stimulator for the neuron, and a transducer for detecting a signal produced in the neuron. The neuronal cell is positioned on the substrate in a predefined orientation. The neuronal cell is conveniently oriented by providing a patterned self-assembled monolayer on the substrate. In a preferred embodiment, a capacitative transducer serves as the transducer and is capable of detecting a signal propagating in the neuron. A network of neuroelectric devices that defines a logic device also is contemplated, which includes a plurality of neurons each in electrical communication with another neuron. The neurons are provided in synaptic relationship so that a signal propagating in one neuron can be attenuated upon stimulation of another neuron.
摘要翻译：神经电学装置由其上提供有神经元细胞的底物，用于神经元的刺激器和用于检测在神经元中产生的信号的换能器来限定。神经元细胞以预定义的方向定位在基底上。通过在衬底上提供图案化的自组装单层，方便地定向神经元细胞。在优选实施例中，电容换能器用作换能器并且能够检测在神经元中传播的信号。还设想了定义逻辑装置的神经电装置的网络，其包括多个与另一个神经元电连通的神经元。神经元以突触关系提供，使得在一个神经元中传播的信号可以在刺激另一个神经元时减弱。

10. 发明申请

US20120122728A1 Method Of Screening Drugs For Reversal Of Amyloid Beta Neurotoxicity 审中-公开
标题翻译：筛选药物用于反转淀粉样蛋白β神经毒性的方法
公开(公告)号：US20120122728A1
公开(公告)日：2012-05-17
申请号：US13322911
申请日：2010-05-27
申请人： James J. Hickman , Kucku Vaghese , Peter Molnar
发明人： James J. Hickman , Kucku Vaghese , Peter Molnar
IPC分类号： C40B30/06 , C12Q1/02
CPC分类号： C07K14/4711
摘要： A method of screening a compound for effectiveness in treating amyloid beta neurotoxicity comprises culturing mammalian neurons in serum-free defined medium until the neurons are electrically functional, exposing the electrically stable neurons to amyloid beta, monitoring the exposed neurons for impairment of electrical functionality, and treating the exposed neurons with the candidate drug while monitoring their electrical activity for reversal of impairment. The invention also includes a method of identifying a mammalian neuron having a biological marker conferring predisposition to development of Alzheimer's disease, the method comprising culturing the mammalian neuron in serum-free medium until the neuron is electrically functional, exposing the electrically stable neuron to amyloid beta while monitoring for impairment of electrical functionality as an indicator of presence of said biological marker, and verifying presence of the biological marker by treating the impaired neuron with an anti-amyloidogenic compound while monitoring for return of neuron functionality.
摘要翻译：筛选化合物以治疗淀粉样蛋白β神经毒性的有效性的方法包括在无血清定义的培养基中培养哺乳动物神经元，直到神经元具有电功能，将电稳定的神经元暴露于淀粉样蛋白β，监测暴露的神经元以损害电功能，以及用候选药物治疗暴露的神经元，同时监测其电活动以逆转损伤。本发明还包括鉴定具有赋予阿尔茨海默病发展倾向的生物学标记的哺乳动物神经元的方法，所述方法包括在无血清培养基中培养哺乳动物神经元，直到神经元具有电功能，将电稳定的神经元暴露于淀粉样蛋白β 同时监测电功能损伤作为所述生物标志物存在的指标，以及通过用抗淀粉样变性化合物治疗受损神经元同时监测神经元功能的返回来验证生物标志物的存在。

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