CN1275885A - 测定包括化学治疗剂在内的活性剂精确效力的方法 - Google Patents

测定包括化学治疗剂在内的活性剂精确效力的方法 Download PDF

Info

Publication number
CN1275885A
CN1275885A CN97196143A CN97196143A CN1275885A CN 1275885 A CN1275885 A CN 1275885A CN 97196143 A CN97196143 A CN 97196143A CN 97196143 A CN97196143 A CN 97196143A CN 1275885 A CN1275885 A CN 1275885A
Authority
CN
China
Prior art keywords
cell
patient
agent
activating agent
sample
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN97196143A
Other languages
English (en)
Other versions
CN1143618C (zh
Inventor
P·L·科恩布里斯
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Precision Therapeutics Inc
Original Assignee
Precision Therapeutics Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Precision Therapeutics Inc filed Critical Precision Therapeutics Inc
Publication of CN1275885A publication Critical patent/CN1275885A/zh
Application granted granted Critical
Publication of CN1143618C publication Critical patent/CN1143618C/zh
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5011—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing antineoplastic activity
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5014—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing toxicity
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis

Abstract

一种用来为具体患者筛选多个候选治疗剂或化学治疗剂的效力的改进系统,其中,先获取患者的组织样品,培育并使其分别暴露在多种处理剂和/或治疗剂下,以便客观地确定对特定患者的最佳治疗剂。具体的方法革新,如组织样品制备方法,使得该方法在实践和理论上均适用。对于初始组织培养单层制备来说,特别重要的组织样品制备方法是初步制备组织样品的粘合性多细胞颗粒,而不是酶解离的细胞悬液或制备液。通过使均匀的细胞样品受各种活性剂(及其各种浓度)的作用,就可确定治疗特定患者最为有效的试剂和浓度。

Description

测定包括化学治疗剂在内的活性剂精确效力的方法
                 发明领域
本发明涉及包括化学治疗剂在内的活性剂的筛选和测试方法，以预测其在打算使用这种试剂来治疗的各患者中的潜在效力。
                   引言
在美国，包括化学治疗剂再内的所有活性剂都应在批准用于医疗前进行效力和安全性的严格测试。评价效力的方法包括对给定治疗方法和/或活性剂用双盲试验(double blind study)在大量试验者中进行细致的研究，然后对获得的数据作统计学整理分析，但是这些结论必定是根据患者总数作为一个整体来作出的。然而，在许多药物学上，尤其在化学治疗领域上，单个患者治疗的结果可能与总结的数据并不相符，这对单个患者是不利的。长期以来认识到需要有这样一种方法，该方法能在对给定的各个患者治疗前测定治疗活性剂(包括但不局限于化学治疗剂)对该患者的治疗潜力。
已有技术测定方法早已存在，该方法是将各类恶性组织暴露在多种活性剂下以评价治疗给药的最佳选择。例如，在Kruczynski，A.等，″Evidence of a directrelationship between the increase in the in vitro passage number of human non-small-cell-lung cancer primocultures and their chemosensitivity，″Anticancer Research，vol.13，no.2，pp.507-513(1993)中，在体内移植物、体外初始培养物(primoculture)和商业上购得的长期生长癌细胞系中，研究了非肺癌小细胞(non-small-cell-lungcancer)的化学敏感性。记录下化学敏感性的增加，并使其与出现问题的细胞的形态变化关联。有时用预期的治疗剂来测试动物模型恶性细胞和/或建立的细胞培养物，例如参见Arnold，J.T.，″Evaluation of chemopreventive agents in differentmechanistic classes using a rat tracheal epithelial cell culture transformation assay，″Cancer Res.，vol.55，no.3，pp.537-543(1995)。
当针对各个患者用真正的患者细胞来进行体外测定方法时，在典型的已有技术方法中，细胞通过获取(活体检查)和胰蛋白酶消化(用胰蛋白酶消化结缔组织)来产生适于变成所需组织培养物形式的细胞悬液。从这些方法获得的体外组织培养细胞收集物通常受到这样的困扰，即它们不能准确地模拟原来的肿瘤或其它活体检查细胞的化学敏感性。而且，标准的克隆和组织培育技术在用于不同患者的测定设置时显得非常复杂和昂贵。因此仍需要一种组织培育方法，该方法能为药物筛选提供细胞培养物，该细胞培养物在简单地制备后能以等价于其体内反应性的方式进行反应，从而能对打算进行这种筛选的特定患者进行药物或化学治疗剂的筛选。
                             发明概述
为了满足这一需求，本发明是用来为具体患者筛选多个候选治疗剂或化学治疗剂的效力的改进系统，其中，先获取患者的组织样品，培育并使其分别暴露在多种处理剂和/或治疗剂下，以客观地确定对患者的培育细胞的最佳处理剂。具体的方法革新，如组织样品制备方法，使得该方法在实践和理论上均适用。对于制备最初的组织培养单层来说，一个特别重要的组织样品制备方法是初步制备组织样品的粘合性多细胞颗粒，而不是酶解离的细胞悬液或制备液。例如对于恶性细胞的培育，认为(不拟受理论束缚)通过将恶性细胞保持在原来组织的多细胞颗粒内，恶性细胞本身的生长比成纤维细胞或其它细胞的过分生长(当悬浮的肿瘤细胞在培养物中生长时很容易发生这种情况)更容易。因此可以形成实用的单层细胞，以便能有意义地筛选多种处理剂和/或试剂。检测细胞的生长以确定开始试验的时间，并测定培育细胞的生长速度；另外对药物加入的次序和时间也进行监控和优化。通过使均匀的细胞样品受各种活性剂(及其各种浓度)的作用，就可确定最有效的试剂。对于有关癌症治疗的测定，考虑采用两阶段评价方法，在该方法中对给定的抗癌试剂的急性细胞毒性和长期抑制效果均可进行研究。
                           发明详述
本发明是用来为具体患者筛选多种候选治疗剂或化学治疗剂的效力的系统，其中，先获取患者的组织样品，并使其分别暴露在多种处理剂和/或治疗剂下，以客观地确定最佳的处剂理或试剂。具体的方法革新，如组织样品制备方法，使得该方法在实践和理论上均适用。对于制备最初的组织培养单层来说，一个特别重要的组织样品制备技术是初步制备组织样品的粘合性多细胞颗粒(外值体)，而不是酶解离的细胞悬液或制备液。对细胞生长、药物加入的次序和时间进行监控和优化。
本发明的一个重要应用是筛选出抗恶性细胞(该恶性细胞从患者体内活体检查获得)组织培养制备物的化学治疗剂和其它抗赘生治疗剂。可用本发明系统来筛选的有关抗癌症治疗剂是，放射治疗剂和增强放射致细胞毒性的试剂，以及抗癌症免疫治疗试剂。非恶性综合征的处理剂或治疗剂的筛选方法也包括在本发明范围内，然而，它还包括抗过度增生综合征(如牛皮癣)的试剂，或伤口愈合剂。本发明的效力测定法不仅仅局限于筛选可加速细胞生长(愈合情况)或减缓细胞生长(抗癌症、抗过度增生)的活性剂，因为本发明的组织培育系统也可用于测试增强或抑制细胞内生物化学功能的试剂。例如，在对患者进行治疗之前，可用本发明的测定方法来筛选酶、神经传递介质和其它生物化学物质的形成或阻断。
在治疗患者肿瘤时，在本发明的较佳实例中，采用本领域已知的任何合适的活体检查或外科方法，来从患者体内获得肿瘤活体检查物，即大于100mg的无坏死、无污染组织。活体检查样品制备通常在层流罩超净台(应在使用前至少20分钟前打开)上按照以下步骤进行。在开始制备样品前，用试剂级乙醇擦干净超净台表面。然后在无菌条件下，从运输容器中取出肿瘤，并用消毒剪剪碎。如果获得的样品已经粉碎，则应将各肿瘤部分分成四组。然后用消毒镊子将每一未分组的组织的1/4部分放入3ml消毒生长培养基(标准的F-10培养基，含有17％小牛血清和标准量的青霉素和链霉素)中，并用两把类似剪刀运动方式的消毒手术刀、或机械上等效的手工或自动的相反切齿刀片(incisor blade)来进行有规则的切削。这种交错切割的运动很重要，因为该方法在获得的肿瘤多细胞颗粒上产生了平滑的切割边缘。较佳的(但不是必需的)，每个肿瘤颗粒的大小为1mm3。在每个肿瘤的1/4部分被切碎后，用消毒的巴斯德毛细吸管将颗粒种在培育瓶中(每个T-25瓶中有9个外值体，或每个T-75瓶中有20个颗粒)。然后在每个瓶上标上患者代码、外植日期以及任何其它辨别的数据。外值体应均匀地分布在瓶底表面上，先在37℃培养箱中倒置培育5-10分钟，然后加入约5-10ml消毒的生长培养基，以正常的非倒置位置进行进一步培养。此瓶放在35℃、没有CO2的培养箱中。应每天检查瓶中的生长和污染情况。在几个星期后(每周取出并更换5ml生长培养基)，外值体会使细胞生长形成单层。对于恶性细胞的培养，认为(不拟受理论束缚)通过将恶性细胞保持在原来组织的多细胞颗粒内，恶性细胞本身的生长比成纤维细胞(或其它不要的细胞)的过度生长(悬浮的肿瘤细胞在培养基中生长时经常产生此情况)更容易。
用上述步骤来形成细胞单层培养物，可以使来自组织样品的恶性细胞的生长最大化，因而优化了随后进行的各种待测试剂化学治疗作用的组织培养测定。然而，真正的恶性细胞生长增强只是本发明的一个方面；本发明的另一重要特征是用来监视一旦形成的单层的生长情况的生长速度监控系统。一旦初始培养及其衍生的次级单层组织培养开始，就对细胞的生长进行监测，以确定开始化学治疗试验的时间并测定培育细胞的生长速度。
对细胞生长的监控这样来进行：不杀死细胞或不对细胞染色，也不从培养瓶中取出任何细胞，对单层细胞进行定期计数。可用肉眼观察或自动的方法，采用或不采用本领域已知的估计方法(例如，计数网格的代表性区域内的数量，再乘以网格数)来进行计数。然后用定期计数获得的数据来确定生长速度是否与同类细胞在患者体内的生长速度平行。例如，如果记录了生长速度周期，则可以为患者制定某些活性剂的剂量。相同的生长速度也可用来评价放射治疗的周期性。应当注意，当单层在中生长时进行生长速度测定时，本发明的方法不需用血细胞计数、流式细胞计数或用显微镜载玻片和染色，也没有这些方法附加的劳力和成本。
细胞单层生长速度的步骤通常是：用相差倒置显微镜检测在37℃培养箱(5％CO2)中培育的培养瓶。将瓶放在相差倒置显微镜下，用10X物镜检测10个网格区域(瓶上固有一个网格区域)，其条件是10个区域不应相邻，也不相隔非常远，这样10个区域是整个瓶的代表性取样。记录每个区域被的细胞占据的百分数，对这些百分数取平均值，然后评价细胞培养物中总的铺满百分数(percentconfluency)。当患者样品被分到两个或三个或多个瓶中时，应计算总的患者样品的平均细胞计数值。计算得到的平均铺满百分数应输入工序记录中，以便能汇编这些数据，并绘制时间-生长曲线。可对单层培养物进行照相，以记录细胞形态和培养物生长模式。适用的公式如下：
             细胞占据面积的估计值铺满百分数＝       观察区域的总面积
因此，例如，当细胞占据的面积的估计值为30％而总区域的面积为100％时，铺满百分数为30/100，或30。
采纳上述步骤可直接用于非肿瘤细胞，它大致上形成了等价的步骤。
采用培育细胞的活性剂筛选不在原先的培育瓶中进行，而通常在板(如微量滴定板)中进行。用于筛选目的的化学敏感性测定方法的实施取决于将可再现细胞数量输递到一块板和/或一系列板每行中的能力，以及在给定孔中使细胞平均分布的能力。下列步骤确保了细胞能再现地从瓶转移到微量滴定板中，且细胞均匀地分布在每个孔的表面上。
制备微量滴定板的第一步当然是如上所述的制备和检测细胞单层。下列步骤是示范性的，对其所作的变动对于本领域技术人员来说是显然的。从培育瓶中取出细胞，通过离心制备细胞沉淀。然后将从单层获得的细胞沉淀悬浮在5ml生长培养基中，并用涡流搅拌器在锥形试管中混合6-10秒。然后来回振荡试管10次。从锥形试管中心吸取36μl的液滴，滴在96孔板的一个孔中。然后用一新的移液管吸取36μl等份的台盼蓝溶液加入同一孔中，靠移液管的反复抽吸来使两液滴混合。然后将获得的混合物分到血细胞计数计的两个小室中，用标准的光学显微镜检测。在10x放大倍数下，对血细胞计数计的四个象限中的两个进行计数。只对那些没有接受台盼蓝染料的细胞进行计数。对第二个计数小室重复该过程。这样就测得了每个室的平均细胞计数。用本领域已知的方法核定象限计数值，作记录(log)，乘以10-4，给出细胞数/毫升，并相应地计算出使其余细胞等份悬浮必需的液体(生长培养基)总量。
在测定了培养基中所需的细胞浓度后，用涡流搅拌机和振荡使来自单层的其余细胞等份悬浮在生长培养基中，并装入本领域已知的Terasaki分装器中。用本领域已知的Terasaki分装技术将制得的细胞悬液按等份转移到微量滴定板上。根据需要可以从一个细胞悬液制得多个培养板。然后根据本领域已知的方法，用消毒的湿棉纱布包裹平板，并放在培养箱中培育。
在制得微量滴定板后，根据下述示范性的步骤使其中的细胞暴露在活性剂下。在本发明测定方法的这部分中，将合适量的特定活性剂转移到如上所述制得的微量滴定板中。通用步骤可作如下改变。解开每个微量滴定板的湿棉纱布，并用显微镜检测细胞的粘附情况。将对照溶液分散在微量滴定板网格中标记行的孔中，在其余行的其余孔中加入合适等份的待测试活性剂。通常是，依次将浓度渐增的待测试活性剂加入培养板中数字逐渐增高的行中。然后将培养板重新包在纱布内，并在37℃、5％CO2的培养箱中培养。在暴露了预定的时间后，解开包布取出培养板，用消毒纱布吸除试剂，用Hank′s Balance Salt Solution洗涤，注入生长培养基，并再次在培养箱中培养预定的时间，然后固定培养板和染色，并进行评价。
固定和染色可根据许多合适的步骤来进行；下面是典型的步骤。从培养箱中取出培养板，倒去培养基，用Hank′s Balance Salt Solution注满培养板。在重复注满倒去后(每次伴随搅拌)，注入试剂级乙醇保持2-5分钟。然后倒去乙醇。每个培养板用约5ml Giemsa Stain液进行染色，但是体积并不是关键，注满才是目的。Giemsa染色应放置5分钟±30秒，因为时间长短会影响染色强度。然后倒去Giemsa染色液，将板浸泡在烧杯中的3倍冷自来水中。然后，翻转平板用力振动，并在实验室工作台支架上空气干燥过夜(除去平板盖子)。然后人工或用自动和/或计算机方法对每个孔中的细胞计数，以获得暴露在不同浓度活性剂下时细胞的化学敏感性数据。一种特别适用于细胞计数的计算机操作环境是商业上购得的OPTIMATE编译程序，该编译程序被设计成能执行光学计数功能，该功能非常适用于计算机化细胞计数步骤以及随后的计算。
当测定细胞生长促进剂而不是细胞抑制剂(如化学治疗剂)时，上述步骤没有作明显改变。本发明测定方法使细胞死亡或细胞生长能同样容易地被检测。在任何一种情况下，本发明系统应用的优化包括各种候选活性剂的对比测试，从而根据体外测试结果为患者选择最佳的治疗候选物。上述发明的一个特别有利的实例包括一个两阶段的测定，在测定细胞毒性后再评价药物的长期抑制效果。因此，可以分开评价化学治疗剂的直接化学治疗效果及其较长期的持续效力。
一种或多种活性剂或化学治疗剂的鉴定在本发明的范围内，其目的是为给定患者筛选任何或所有试剂的效力。几乎所有的活性剂均可用本发明的方法来进行筛选；因此这里省略了典型活性剂的一览表。
因此，本发明的精髓包括本发明系统重要特征的简单性：用待测试患者组织的粘合性多细胞颗粒来形成细胞单层；对那些单层的生长进行监控以精确地预测相同细胞在体内的相关生长；可对多种活性剂的不同浓度进行测试，目的不但是确定最合适的试剂，而且是确定用于真实患者治疗时该试剂的最佳浓度(根据计算的细胞生长速度)。也应注意在本发明的内容中，本发明系统允许在组织培养单层(该培养物在快速生长条件下(约几周)在营养培养基中生长)的悬液中进行体外测试，而不是用稀释克隆经过长时间产生的单一细胞子代来测试。在一些情况下，本发明是测定细胞毒性和长期生长抑制作用的两阶段测定法。
尽管本发明已就上述特定材料和方法进行了描述，但应认为，本发明只局限在附加的权利要求提出的范围内。

Claims (12)

1.一种评价患者细胞化学敏感性的方法，该方法包括下列步骤：
a)收获患者组织、细胞腹水或渗出液的样品；
b)将所述样品分成多细胞颗粒；
c)从所述粘合性多细胞颗粒中长出组织培养单层；
d)将所述单层的细胞接种入分隔的多个部位；和
e)用至少一种活性剂处理所述多个部位，然后评价所述部位中的细胞对至少一种活性剂的化学敏感性。
2.根据权利要求1所述的方法，其中步骤a)还包括步骤
a)制备一个样品，该样品从患者肿瘤组织的样品中获得。
3.根据权利要求1所述的方法，其中所述分隔的多个部位还包括其中含有多个孔的平板。
4.根据权利要求1所述的方法，其中步骤e)还包括步骤
e)用不同浓度的多种活性剂来处理所述多个部位，然后评价单种活性剂在单个浓度下的最优化学敏感性。
5.根据权利要求1所述的方法，其中步骤e)还包括步骤
e)用多种活性剂对所述多个部位处理充分长的时间，以评价所述多种活性剂的至少一种的初始细胞毒性效果和长期抑制效果。
6.根据权利要求1所述的方法，其中根据步骤e)测得的化学敏感性是抗癌敏感性。
7.根据权利要求1所述的方法，其中步骤d)用Terasaki分装器来实现。
8.根据权利要求1所述的方法，其中在步骤d)中，细胞在接种入培养板的多个孔中之前，先制备成悬液。
9.根据权利要求1所述的方法，其中所述活性剂是化学治疗剂。
10.根据权利要求1所述的方法，其中所述活性剂是伤口愈合剂。
11.根据权利要求1所述的方法，其中所述活性剂是放射治疗剂和/或放射治疗致敏剂或改进剂。
12.根据权利要求1所述的方法，其中所述活性剂是免疫治疗剂。
CNB971961433A 1996-07-12 1997-07-10 测定包括化学治疗剂在内的活性剂精确效力的方法 Expired - Fee Related CN1143618C (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US08/679,056 US5728541A (en) 1996-07-12 1996-07-12 Method for preparing cell cultures from biologial specimens for chemotherapeutic and other assays
US08/679,056 1996-07-12

Publications (2)

Publication Number Publication Date
CN1275885A true CN1275885A (zh) 2000-12-06
CN1143618C CN1143618C (zh) 2004-03-31

Family

ID=24725407

Family Applications (1)

Application Number Title Priority Date Filing Date
CNB971961433A Expired - Fee Related CN1143618C (zh) 1996-07-12 1997-07-10 测定包括化学治疗剂在内的活性剂精确效力的方法

Country Status (13)

Country Link
US (13) US5728541A (zh)
EP (1) EP0912085B1 (zh)
JP (1) JP4108128B2 (zh)
CN (1) CN1143618C (zh)
AT (1) ATE360989T1 (zh)
AU (1) AU712302B2 (zh)
BR (1) BR9710348A (zh)
CA (1) CA2259984C (zh)
DE (1) DE69737684T2 (zh)
ES (1) ES2283022T3 (zh)
IL (1) IL127964A (zh)
NZ (1) NZ333830A (zh)
WO (1) WO1998002038A1 (zh)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105143883A (zh) * 2013-04-25 2015-12-09 Q吉尔股份有限公司 基于细胞的药物筛选分析方法和其用途
CN105263578A (zh) * 2013-03-28 2016-01-20 蒙彼利埃大学医疗中心 确定放射敏感性的方法
CN116622801A (zh) * 2023-07-24 2023-08-22 安泰康生物技术(北京)有限公司 一种患者来源超微组织培养药物敏感性检测方法

Families Citing this family (67)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040072722A1 (en) * 2002-10-10 2004-04-15 Kornblith Paul L. Methods for assessing efficacy of chemotherapeutic agents
US20040023375A1 (en) * 2002-07-30 2004-02-05 Precision Therapeutics, Inc. Method for preparing cell cultures from biological specimens for chemotherapeutic and other assays
US6416967B2 (en) * 1996-07-12 2002-07-09 Precision Therapeutics, Inc. Method of using multicellular particulates to analyze malignant or hyperproliferative tissue
US5728541A (en) * 1996-07-12 1998-03-17 Precision Therapeutics, Inc. Method for preparing cell cultures from biologial specimens for chemotherapeutic and other assays
US6171780B1 (en) 1997-06-02 2001-01-09 Aurora Biosciences Corporation Low fluorescence assay platforms and related methods for drug discovery
US6517781B1 (en) * 1997-06-02 2003-02-11 Aurora Biosciences Corporation Low fluorescence assay platforms and related methods for drug discovery
US6861035B2 (en) * 1998-02-24 2005-03-01 Aurora Discovery, Inc. Multi-well platforms, caddies, lids and combinations thereof
DE19912798C1 (de) * 1999-03-10 2000-02-17 Andreas Jordan Verfahren zur Kultivierung von Krebszellen aus Humangewebe und Vorrichtung zur Aufbereitung von Gewebeproben
US20040086888A1 (en) * 2001-10-18 2004-05-06 Kornblith Paul L Method for tandem genomic/proteomic analysis of proliferating cells
US7276351B2 (en) 2003-09-10 2007-10-02 Seahorse Bioscience Method and device for measuring multiple physiological properties of cells
US8658349B2 (en) 2006-07-13 2014-02-25 Seahorse Bioscience Cell analysis apparatus and method
US20070087401A1 (en) * 2003-10-17 2007-04-19 Andy Neilson Analysis of metabolic activity in cells using extracellular flux rate measurements
US20080064034A1 (en) * 2004-05-21 2008-03-13 Fraunhofer-Gesellschaft Zur Förderung Der Angewand Multi-Cellular Test Systems
AU2006287175A1 (en) * 2005-09-01 2007-03-08 Precision Therapeutics, Inc. Chemo-sensitivity assays using tumor cells exhibiting persistent phenotypic characteristics
EP2136630A4 (en) * 2007-03-23 2010-06-02 Precision Therapeutics Inc METHODS FOR ASSESSING ANGIOGENIC POTENTIAL IN CULTURE
WO2008150496A2 (en) * 2007-05-31 2008-12-11 Genelux Corporation Assay for sensitivity to chemotherapeutic agents
EP2207892A4 (en) * 2007-10-15 2010-12-08 Precision Therapeutics Inc METHOD FOR SELECTION OF ACTIVE AGENTS FOR CANCER TREATMENT
US20090136917A1 (en) * 2007-10-25 2009-05-28 Szalay Aladar A Systems and methods for viral therapy
ES2545760T3 (es) 2008-02-25 2015-09-15 Nestec S.A. Selección de fármacos para terapia del cáncer de mama utilizando matrices de anticuerpos
US20100311084A1 (en) * 2008-05-14 2010-12-09 Precision Therapeutics, Inc. Methods for predicting a patient's response to egfr inhibitors
EP2281027A4 (en) * 2008-05-14 2011-07-27 Precision Therapeutics Inc METHOD FOR PREDICTING THE REACTION OF A PATIENT TO EGFR HEMMER
WO2010003057A2 (en) 2008-07-03 2010-01-07 Mayo Foundation For Medical Education And Research Treating cancer
US8202702B2 (en) * 2008-10-14 2012-06-19 Seahorse Bioscience Method and device for measuring extracellular acidification and oxygen consumption rate with higher precision
US20110238322A1 (en) * 2008-11-03 2011-09-29 Precision Therapeutics, Inc. Methods of simulating chemotherapy for a patient
US8357175B2 (en) * 2008-12-16 2013-01-22 Nico Corporation Positioning system for tissue removal device
US9216031B2 (en) * 2008-12-16 2015-12-22 Nico Corporation Tissue removal device with adjustable fluid supply sleeve for neurosurgical and spinal surgery applications
US20100152762A1 (en) * 2008-12-16 2010-06-17 Mark Joseph L Tissue removal system with multi-directional foot actuator assembly for neurosurgical and spinal surgery applications
US8496599B2 (en) * 2008-12-16 2013-07-30 Nico Corporation Tissue removal device for neurosurgical and spinal surgery applications
US10368890B2 (en) 2008-12-16 2019-08-06 Nico Corporation Multi-functional surgical device for neurosurgical and spinal surgery applications
US9820480B2 (en) 2008-12-16 2017-11-21 Nico Corporation System for collecting and preserving tissue cores
US8702738B2 (en) * 2008-12-16 2014-04-22 Nico Corporation Tissue removal device for neurosurgical and spinal surgery applications
US9504247B2 (en) 2008-12-16 2016-11-29 Nico Corporation System for collecting and preserving tissue cores
US9655639B2 (en) * 2008-12-16 2017-05-23 Nico Corporation Tissue removal device for use with imaging devices in neurosurgical and spinal surgery applications
US8657841B2 (en) * 2008-12-16 2014-02-25 Nico Corporation Tissue removal device for neurosurgical and spinal surgery applications
US8460327B2 (en) * 2008-12-16 2013-06-11 Nico Corporation Tissue removal device for neurosurgical and spinal surgery applications
US9279751B2 (en) 2008-12-16 2016-03-08 Nico Corporation System and method of taking and collecting tissue cores for treatment
US9931105B2 (en) 2008-12-16 2018-04-03 Nico Corporation System and method of taking and collecting tissue cores for treatment
US10080578B2 (en) 2008-12-16 2018-09-25 Nico Corporation Tissue removal device with adjustable delivery sleeve for neurosurgical and spinal surgery applications
US8430825B2 (en) * 2008-12-16 2013-04-30 Nico Corporation Tissue removal device for neurosurgical and spinal surgery applications
WO2011005504A1 (en) 2009-06-22 2011-01-13 Precision Therapeutics, Inc. Methods for predicting a cancer patient's response to antifolate therapy
AU2010273319B2 (en) 2009-07-15 2015-01-22 Nestec S.A. Drug selection for gastric cancer therapy using antibody-based arrays
US20110097748A1 (en) * 2009-10-28 2011-04-28 Warsaw Orthopedic, Inc. Use of cell lines to determine levels of efficacy of pharmaceutical formulations
JP5847733B2 (ja) 2010-01-28 2016-01-27 ザ リージェンツ オブ ザ ユニバーシティ オブ ミシガン 懸滴用アレイプレート
SI2707030T1 (sl) 2011-05-09 2020-10-30 Mayo Foundation For Medical Education And Research Zdravljenje raka
WO2013036753A1 (en) * 2011-09-07 2013-03-14 The Scripps Research Institute Chiral compounds of varying conformational rigidity and methods of synthesis
EP2804538B1 (en) 2012-01-17 2018-03-28 Nico Corporation System for collecting and preserving tissue cores
EP2870480A4 (en) * 2012-07-05 2016-03-23 Caldera Health Ltd METHOD FOR DETERMINING PERSONALIZED TREATMENT COMPOSITIONS FOR PROSTATE CANCER AND BREAST CANCER
EP3967306A1 (en) 2012-10-01 2022-03-16 Mayo Foundation for Medical Education and Research Cancer treatments
US9494577B2 (en) 2012-11-13 2016-11-15 Seahorse Biosciences Apparatus and methods for three-dimensional tissue measurements based on controlled media flow
DE102013010528B3 (de) * 2013-06-25 2014-02-20 FLACOD GmbH Verfahren zur Erstellung einer Datenbank für die Vorabschätzung der Wirksamkeit von Wirkstoffen in der Tumortherapie
EP3148700B1 (en) 2014-06-02 2020-03-18 Agilent Technologies, Inc. Single column microplate system and carrier for analysis of biological samples
WO2015195476A1 (en) 2014-06-16 2015-12-23 Mayo Foundation For Medical Education And Research Treating myelomas
EP3158487A1 (en) * 2014-06-20 2017-04-26 Connecticut Children's Medical Center Automated cell culture system and corresponding methods
US9446148B2 (en) 2014-10-06 2016-09-20 Mayo Foundation For Medical Education And Research Carrier-antibody compositions and methods of making and using the same
TW201707725A (zh) 2015-08-18 2017-03-01 美國馬友醫藥教育研究基金會 載體－抗體組合物及其製造及使用方法
TW201713360A (en) 2015-10-06 2017-04-16 Mayo Foundation Methods of treating cancer using compositions of antibodies and carrier proteins
EP3399861A4 (en) 2016-01-07 2019-08-07 Mayo Foundation for Medical Education and Research INTERFERON CANCER TREATMENT METHODS
US11351254B2 (en) 2016-02-12 2022-06-07 Mayo Foundation For Medical Education And Research Hematologic cancer treatments
CA3018340A1 (en) 2016-03-21 2017-09-28 Mayo Foundation For Medical Education And Research Methods for improving the therapeutic index for a chemotherapeutic drug
EP3432926A4 (en) 2016-03-21 2019-11-20 Mayo Foundation for Medical Education and Research METHODS OF REDUCING THE TOXICITY OF A CHEMOTHERAPEUTIC MEDICAMENT
US10618969B2 (en) 2016-04-06 2020-04-14 Mayo Foundation For Medical Education And Research Carrier-binding agent compositions and methods of making and using the same
JP2019526579A (ja) 2016-09-01 2019-09-19 マヨ ファウンデーション フォー メディカル エデュケーション アンド リサーチＭａｙｏ Ｆｏｕｎｄａｔｉｏｎ Ｆｏｒ Ｍｅｄｉｃａｌ Ｅｄｕｃａｔｉｏｎ Ａｎｄ Ｒｅｓｅａｒｃｈ Ｔ細胞癌を標的とする為の方法及び組成物
KR20220151022A (ko) 2016-09-01 2022-11-11 메이오 파운데이션 포 메디칼 에쥬케이션 앤드 리써치 암 치료용 담체－ｐｄ－ｌ１ 결합제 조성물
JP7025412B2 (ja) 2016-09-06 2022-02-24 マヨ ファウンデーション フォー メディカル エデュケーション アンド リサーチ パクリタキセル－アルブミン－結合剤組成物並びに該組成物の使用及び製造方法
US11590098B2 (en) 2016-09-06 2023-02-28 Mayo Foundation For Medical Education And Research Methods of treating triple-negative breast cancer using compositions of antibodies and carrier proteins
US11427637B2 (en) 2016-09-06 2022-08-30 Mayo Foundation For Medical Education And Research Methods of treating PD-L1 expressing cancer
US20190168213A1 (en) * 2017-11-10 2019-06-06 Reliant Immune Diagnostics, Inc. System and method for determining efficacy and dosage using parallel/serial dual microfluidic chip

Family Cites Families (44)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4423145A (en) * 1981-05-07 1983-12-27 Stampfer Martha R Enhanced growth medium and method for culturing human mammary epithelial cells
US4668618A (en) 1981-09-14 1987-05-26 Thornthwaite Jerry T Nuclear isolation medium and procedure for separating cell nuclei
US4937187A (en) * 1983-02-04 1990-06-26 Brown University Research Foundation Methods for separating malignant cells from clinical specimens
US4559299A (en) 1983-02-04 1985-12-17 Brown University Research Foundation Inc. Cytotoxicity assays in cell culturing devices
US4937182A (en) 1985-12-19 1990-06-26 Peralta Cancer Research Institute Method for predicting chemosensitivity of anti-cancer drugs
US4816395A (en) 1985-12-19 1989-03-28 Peralta Cancer Research Institute Method for predicting chemosensitivity of anti-cancer drugs
US5266480A (en) * 1986-04-18 1993-11-30 Advanced Tissue Sciences, Inc. Three-dimensional skin culture system
US4996145A (en) 1987-10-27 1991-02-26 Oncotech Incorporated Method for detecting immune-mediated cytotoxicity
US5258541A (en) * 1988-02-16 1993-11-02 Daicel Chemical Industries, Ltd. Process for producing carbonic acid ester
US5242806A (en) * 1990-05-07 1993-09-07 Baxter Diagnostics Inc. Method for conducting the cytotoxicity assays on tumor cells
US5874218A (en) 1990-06-11 1999-02-23 Nexstar Pharmaceuticals, Inc. Method for detecting a target compound in a substance using a nucleic acid ligand
US5747469A (en) 1991-03-06 1998-05-05 Board Of Regents, The University Of Texas System Methods and compositions comprising DNA damaging agents and p53
US5270172A (en) * 1991-04-26 1993-12-14 Dekk-Tek, Inc. Method to predict tumor response to therapy
US5407653A (en) 1991-06-26 1995-04-18 Brigham And Women's Hospital Evaluation of the multidrug resistance phenotype
JPH07503943A (ja) 1991-10-29 1995-04-27 クローバー　コンソリデイテッド，リミテッド 封入及び薬剤放出に有用な架橋性の多糖類、ポリカチオン及び脂質
WO1994000470A1 (en) 1992-06-30 1994-01-06 The Wistar Institute Of Anatomy And Biology DEVELOPMENTAL EMBRYONIC MOUSE cDNA LIBRARIES
US7166423B1 (en) * 1992-10-21 2007-01-23 Miltenyi Biotec Gmbh Direct selection of cells by secretion product
US5856112A (en) * 1994-06-16 1999-01-05 Urocor, Inc. Method for selectively inducing biomarker expression in urologic tumor tissue for diagnosis and treatment thereof
GB9419716D0 (en) 1994-09-30 1994-11-16 Gearey David Electrochemical assessment of cell behaviour and metabolic activity
US5607918A (en) 1995-03-01 1997-03-04 Ludwig Institute For Cancer Research Vascular endothelial growth factor-B and DNA coding therefor
US5888765A (en) 1995-06-23 1999-03-30 President And Fellows Of Harvard College Endothelial-cell specific promoter
US6274576B1 (en) 1995-06-27 2001-08-14 The Henry Jackson Foundation For The Advancement Of Military Medicine Method of dynamic retardation of cell cycle kinetics to potentiate cell damage
US6020473A (en) 1995-08-25 2000-02-01 Genentech, Inc. Nucleic acids encoding variants of vascular endothelial cell growth factor
US5942385A (en) 1996-03-21 1999-08-24 Sugen, Inc. Method for molecular diagnosis of tumor angiogenesis and metastasis
US20020168679A1 (en) 1998-06-11 2002-11-14 Gregory J. Naus Staining agents and protocols for characterizing malignant cells in culture
US20040023375A1 (en) * 2002-07-30 2004-02-05 Precision Therapeutics, Inc. Method for preparing cell cultures from biological specimens for chemotherapeutic and other assays
US6416967B2 (en) * 1996-07-12 2002-07-09 Precision Therapeutics, Inc. Method of using multicellular particulates to analyze malignant or hyperproliferative tissue
US20040072722A1 (en) 2002-10-10 2004-04-15 Kornblith Paul L. Methods for assessing efficacy of chemotherapeutic agents
US5728541A (en) * 1996-07-12 1998-03-17 Precision Therapeutics, Inc. Method for preparing cell cultures from biologial specimens for chemotherapeutic and other assays
US6008007A (en) 1997-01-31 1999-12-28 Oncotech, Inc. Radiation resistance assay for predicting treatment response and clinical outcome
US5840507A (en) 1997-03-19 1998-11-24 Oncotech, Inc. Methods for cancer prognosis and diagnosis
US5972639A (en) 1997-07-24 1999-10-26 Irori Fluorescence-based assays for measuring cell proliferation
US6111092A (en) 1998-06-03 2000-08-29 Millennium Pharmaceuticals, Inc. Nucleic acid molecules encoding DRT111 and uses thereof
US6664062B1 (en) 1998-07-20 2003-12-16 Nuvelo, Inc. Thymidylate synthase gene sequence variances having utility in determining the treatment of disease
WO2000052204A2 (en) 1999-02-22 2000-09-08 Orntoft Torben F Gene expression in bladder tumors
AU1465801A (en) 1999-11-03 2001-05-14 Oncotech, Inc. Methods for cancer prognosis and diagnosis
AU2002224435A1 (en) 2000-10-18 2002-04-29 Precision Therapeutics, Inc. Method for tandem genomic/proteomic analyses of proliferating cells
US7244578B2 (en) * 2001-04-06 2007-07-17 Department Of Veterans Affairs Methods for modeling infectious disease and chemosensitivity in cultured cells and tissues
US20040086888A1 (en) 2001-10-18 2004-05-06 Kornblith Paul L Method for tandem genomic/proteomic analysis of proliferating cells
US7314733B2 (en) * 2001-11-13 2008-01-01 Albert Einstein College Of Medicine Of Yeshiva University Saitohin gene and uses of same
US6900017B2 (en) * 2002-01-10 2005-05-31 The Board Of Trustees Of The University Of Illinois Human hematopoietic growth regulatory gene and uses
JP4175215B2 (ja) * 2003-08-08 2008-11-05 日産自動車株式会社 バイポーラ電池、組電池、複合組電池、および組電池または複合組電池を用いた車両
RU2007130552A (ru) * 2005-01-10 2009-02-20 Рисерч Дивелопмент Фаундейшн (US) Рекомбинантные молекулы направленного действия для лечения рака
AU2006287175A1 (en) * 2005-09-01 2007-03-08 Precision Therapeutics, Inc. Chemo-sensitivity assays using tumor cells exhibiting persistent phenotypic characteristics

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105263578A (zh) * 2013-03-28 2016-01-20 蒙彼利埃大学医疗中心 确定放射敏感性的方法
CN105263578B (zh) * 2013-03-28 2018-10-02 蒙彼利埃大学医疗中心 确定放射敏感性的方法
CN105143883A (zh) * 2013-04-25 2015-12-09 Q吉尔股份有限公司 基于细胞的药物筛选分析方法和其用途
CN116622801A (zh) * 2023-07-24 2023-08-22 安泰康生物技术(北京)有限公司 一种患者来源超微组织培养药物敏感性检测方法

Also Published As

Publication number Publication date
AU3649397A (en) 1998-02-09
ATE360989T1 (de) 2007-05-15
US20100120070A1 (en) 2010-05-13
US20080076146A1 (en) 2008-03-27
CA2259984A1 (en) 1998-01-22
US5728541A (en) 1998-03-17
US20050202411A1 (en) 2005-09-15
US20100330613A1 (en) 2010-12-30
CN1143618C (zh) 2004-03-31
US7501260B2 (en) 2009-03-10
US6933129B1 (en) 2005-08-23
DE69737684T2 (de) 2008-01-31
US8058025B2 (en) 2011-11-15
US7112415B2 (en) 2006-09-26
US6900027B1 (en) 2005-05-31
EP0912085A1 (en) 1999-05-06
US8183009B2 (en) 2012-05-22
EP0912085A4 (en) 2002-07-03
IL127964A (en) 2001-07-24
NZ333830A (en) 2002-12-20
US20070037136A1 (en) 2007-02-15
US20090042221A1 (en) 2009-02-12
ES2283022T3 (es) 2007-10-16
JP4108128B2 (ja) 2008-06-25
EP0912085B1 (en) 2007-05-02
CA2259984C (en) 2008-10-07
BR9710348A (pt) 2000-01-11
AU712302B2 (en) 1999-11-04
US6887680B2 (en) 2005-05-03
US20120088264A1 (en) 2012-04-12
US7678552B2 (en) 2010-03-16
WO1998002038A1 (en) 1998-01-22
US7314731B2 (en) 2008-01-01
JP2002501609A (ja) 2002-01-15
DE69737684D1 (de) 2007-06-14
US20050202410A1 (en) 2005-09-15
US20020192638A1 (en) 2002-12-19
US20090042225A1 (en) 2009-02-12
US7771963B2 (en) 2010-08-10
IL127964A0 (en) 1999-11-30
US7563593B2 (en) 2009-07-21

Similar Documents

Publication Publication Date Title
CN1143618C (zh) 测定包括化学治疗剂在内的活性剂精确效力的方法
US7972769B2 (en) Method for preparing cell cultures from biological specimens for chemotherapeutic and other assays
US6416967B2 (en) Method of using multicellular particulates to analyze malignant or hyperproliferative tissue
Norrby In vivo models of angiogenesis
Herlenius et al. Functional stem cell integration assessed by organotypic slice cultures
RU2154270C1 (ru) Способ определения биологической активности препарата
Drew et al. Studies on the survival of algae added to chemically treated soils—1. methodology
Genoni Short-term effect of a toxicant on scope for change in ascendency in a microcosm community
JPWO2020049642A1 (ja) 細胞解析装置及び細胞解析方法

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
C17 Cessation of patent right
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20040331

Termination date: 20130710