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1. 发明授权

US5712161A Method for culturing animal cells in collagen drops on a support 失效
标题翻译：在支架上培养胶原蛋白中的动物细胞的方法
公开(公告)号：US5712161A
公开(公告)日：1998-01-27
申请号：US507418
申请日：1995-08-24
申请人： Masahiro Koezuka , Naohito Kondo , Hisayuki Kobayashi , Hiroshi Saeki , Keizo Tanisaka , Sachiko Oda
发明人： Masahiro Koezuka , Naohito Kondo , Hisayuki Kobayashi , Hiroshi Saeki , Keizo Tanisaka , Sachiko Oda
IPC分类号： C12N5/00 , C12N5/08 , C12N1/02 , C12N11/04 , C12Q1/02
CPC分类号： C12N5/0012 , C12N2533/54
摘要： Animal cells are cultured while embedded in a collagen gel. The gel containing cells is formed by dispersing animal cells in a collagen solution, placing a drop (or drops) of the cell-containing collagen solution on a support surface and allowing the drop to gel to fix on the surface as a globular collagen gel having a convex surface. The cells are cultured by contacting the gel with a culture medium that may be serum-free or contain dextran sulfate. The drop preferably contains about 3 to about 300 microliters of the collagen solution and is about 2 mm or less in height. The cells may be precultured on a support surface having a collagen layer, released from the collagen layer by treatment with collagenase and dispersed in the collagen solution. The cells can be evaluated after culturing by staining such as with neutral red, or with fluorescein diacetate and irradiating, or by photographing cells in the collagen gel. The support surface can be a continuous ridge forming a recessed area and the drop of collagen solution is placed in the recessed area. Cancer cells can be cultured to test for sensitivity to an anticancer agent. The interaction of cells can be observed in co-culture tests where different kinds of cells are cultured simultaneously. Culturing in a drop enables using a small amount of animal cells sampled from tissue.
摘要翻译： PCT No.PCT / JP94 / 02222 Sec。 371日期1995年8月24日 102（e）日期1995年8月24日PCT 1994年12月26日PCT PCT。公开号WO95 / 18216 日期1995年7月6日在植入胶原凝胶的同时培养动物细胞。含有凝胶的细胞通过将动物细胞分散在胶原溶液中，将含有细胞的胶原溶液的滴（或滴）放置在支持体表面上并使滴下凝胶固定在表面上，形成为具有球形胶原凝胶一个凸面。通过使凝胶与可以是无血清或含有葡聚糖硫酸盐的培养基接触来培养细胞。该液滴优选含有约3至约300微升的胶原溶液，并且高度为约2mm或更小。细胞可以在具有胶原层的支撑表面上进行预培养，胶原层通过用胶原酶处理从胶原层释放并分散在胶原溶液中。可以通过染色如中性红培养或用荧光素二乙酸酯进行培养来照射细胞，并照射或通过拍摄胶原凝胶中的细胞。支撑表面可以是形成凹陷区域的连续脊，并且胶原溶液的液滴被放置在凹陷区域中。可以培养癌细胞以测试对抗癌剂的敏感性。在同时培养不同种类的细胞的共培养试验中可观察到细胞的相互作用。在一滴中培养使得能够使用从组织中取样的少量动物细胞。

2. 发明授权

US5356793A Method for testing the sensitivity of anticancer drug 失效
标题翻译：抗癌药物敏感性检测方法
公开(公告)号：US5356793A
公开(公告)日：1994-10-18
申请号：US649526
申请日：1991-02-01
申请人： Masahiro Koezuka , Naohito Kondo , Sachiko Oda , Hisayuki Kobayashi , Masayuki Yasutomi
发明人： Masahiro Koezuka , Naohito Kondo , Sachiko Oda , Hisayuki Kobayashi , Masayuki Yasutomi
IPC分类号： G01N15/14 , G01N33/50 , C12M1/00 , C12M3/00 , C12N5/00 , C12N5/06
CPC分类号： G01N33/5011 , G01N15/1475 , G01N2800/52 , Y10S435/808
摘要： The present invention relates to a method of testing the sensitivity of cancer drugs with cancer cells cultured in vitro. Cancer cells are cultured in a collagen gel substrate. A wide variety of human cancer cell types readily proliferate in the collagen gel substrate, however, fibroblast cells proliferate as well. The measurement of the growth of the cancer cells is hindered by the presence of the fibroblast cells. The present invention solves this problem by counting the number of colonies with an image processor which selectively extracts the image signals of cancer cells and their colonies. In a second embodiment, the growth of cancer cells is determined by measuring the volume of colonies with the image signals of cancer cells and their colonies selectively extracted. The results can be obtained effectively within a short period of time.
摘要翻译：本发明涉及一种检测癌症药物与体外培养的癌细胞的敏感性的方法。癌细胞在胶原凝胶底物中培养。各种人类癌细胞类型在胶原凝胶底物中容易增殖，然而，成纤维细胞也增殖。癌细胞生长的测量被成纤维细胞的存在所阻碍。本发明通过用选择性提取癌细胞及其菌落的图像信号的图像处理器计数菌落数来解决这个问题。在第二个实施方案中，癌细胞的生长通过用选择性提取的癌细胞及其菌落的图像信号测量菌落的体积来确定。结果可以在短时间内有效获得。

3. 发明授权

US4975527A Tissue-affinitive collagen for osteogenesis and method of producing the same 失效
标题翻译：用于成骨的组织亲和胶原及其制备方法
公开(公告)号：US4975527A
公开(公告)日：1990-12-04
申请号：US372115
申请日：1989-06-28
申请人： Masahiro Koezuka , Kunio Takaoka , Kaneo Suzuki , Shigeo Yasugi
发明人： Masahiro Koezuka , Kunio Takaoka , Kaneo Suzuki , Shigeo Yasugi
IPC分类号： A61K35/12 , A61F2/00 , A61K35/32 , A61K35/36 , A61K38/00 , A61K38/17 , A61L27/00 , A61L27/22 , A61L27/24 , C07K14/51 , C07K14/78 , C08H1/06 , C12P21/00
CPC分类号： C07K14/78 , A61L27/227 , A61L27/24 , C07K14/51 , C08H1/06 , A61F2310/00365 , A61K38/00 , Y10S514/801 , Y10S530/84
摘要： A good tissue-affinity and lack of antigenicity are required for the collagen used as the carrier of BMP. Conventional collagens have these properties only insufficiently. When the tyrosine content is below 2 residues/1000 residues, then the above properties of such collagens are good. Moreover, collagens excelling in the above properties can be readily obtained by using proteolytic enzymes.
摘要翻译：对于用作BMP载体的胶原，需要良好的组织亲和性和抗原性不足。常规胶原具有这些性质不足。当酪氨酸含量低于2个残基/ 1000个残基时，这些胶原蛋白的上述性质是好的。此外，通过使用蛋白水解酶可以容易地获得在上述性质中优异的胶原。

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