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公开(公告)号：KR101832133B1
公开(公告)日：2018-02-27
申请号：KR1020110107727
申请日：2011-10-20
Applicant: (주)차바이오텍 , 성균관대학교산학협력단
Abstract: 본발명은환자의폐암조직으로부터효과적인폐암세포의분리및 증식을위한배양조건및 상기의방법으로배양된폐암세포를이용한환자특이적암치료제의스크리닝방법에관한것으로, 보다구체적으로 1) 환자로부터분리한폐암조직을지름 40 내지 100 μm의크기로절개하는단계, 2) 상기 1)단계의절개된조직을배양하는단계, 3) 상기 2)단계의배양된세포에디스파아제를사용하여폐암세포를분리하는단계, 4) 상기 3)단계의폐암세포를코팅물질이코팅된배양접시에서무혈청 N2 배지로배양하여폐암세포만을분리하여배양하는단계, 및 5) 상기 4)단계의분리배양된폐암세포를혈청및 성장인자를첨가한배지에서배양하여증식시키는단계를포함하는폐암조직으로부터폐암세포를분리및 증식시키는방법및 상기방법으로제조된폐암세포를이용하여환자맞춤형암치료제를스크리닝하는방법에관한것이다.
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公开(公告)号：KR1020130043520A
公开(公告)日：2013-04-30
申请号：KR1020110107727
申请日：2011-10-20
Applicant: (주)차바이오텍 , 성균관대학교산학협력단
Abstract: PURPOSE: A method for isolating and proliferating lung cancer cells is provided to effectively isolate and proliferate the lung cancer cells from a patient with lung cancer, and to establish a lung cancer cell line. CONSTITUTION: A method for isolating and proliferating lung cancer cells from a lung cancer tissue comprises: a step of making an incision of a lung cancer tissue from a patient in a diameter of 40-100 micrometers; a step of culturing the lung cancer tissue; a step of isolating lung cancer cells using dispase; a step of culturing the lung cancer cells in a serum-free N2 medium; and a step of culturing the lung cancer cells in a medium containing serum and growth factors.
Abstract translation: 目的：提供肺癌细胞分离和增殖的方法，以有效地从肺癌患者中分离和增殖肺癌细胞，并建立肺癌细胞系。 构成：从肺癌组织分离和增殖肺癌细胞的方法包括：从直径为40-100微米的患者切下肺癌组织的步骤; 培养肺癌组织的一个步骤; 使用分泌酶分离肺癌细胞的步骤; 在无血清N2培养基中培养肺癌细胞的步骤; 以及在含有血清和生长因子的培养基中培养肺癌细胞的步骤。
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公开(公告)号：KR1020140047343A
公开(公告)日：2014-04-22
申请号：KR1020120113492
申请日：2012-10-12
Applicant: (주)차바이오텍
Abstract: The present invention relates to culture conditions for effective isolation and proliferation of lung cancer cells from a lung cancer tissue of a patient, and a method for screening a patient-specific cancer therapeutic agent using the lung cancer cells cultured thereby, and more specifically, to a method for isolating and culturing lung cancer cells from a lung cancer tissue and a method for screening a patient-specific cancer therapeutic agent using the lung cancer cells prepared by the method. The method for isolating and culturing lung cancer cells from a lung cancer tissue, comprises the steps of: 1) cutting a lung cancer tissue isolated from a patient into a size of 40-60 μm in diameter; 2) allowing the cut tissue in step 1) to react with an enzyme to be separated into single cells; 3) proliferating the separated single cells in step 2) in a serum-free ACL4 medium or N2 added medium on a low-adhesion culture dish through suspension culture; and 4) subculturing the cells, which proliferate through the suspension culture in step 3), using Accutase, wherein step 4) does not require a separate Accutase reaction termination step. [Reference numerals] (A) Isolating cells from a cancer tissue according to the conventional art; (AA) Solid tumor; (B) Isolating and culturing lung cancer cells from a lung cancer tissue according to the present invention; (BB) Collagenase degradation; (CC) Single cell suspension; (DD) Cells in agar matrix; (EE) Incubation for 6-8 days; (FF) Cell population formation; (GG) Clonogenic cell isolation; (HH) Isolated clonogenic cell re-dispensing; (II) Cutting into a size of 40-100 um; (JJ) Suspension culture in an ultra-low adhesion (or pluronic F127 coating) culture dish; (KK) Effective cell Formation and culture in a serum-free medium; (LL) Suspension-cultured cell subculture using Accutase; (MM) Adherent culture; (NN) Suspension culture; (OO) EPCAM expression (%); (PP) Adherent culture; (QQ) Suspension culture; (RR) Culture and proliferation through suspension culture for suppressing an EMT phenomenon occurring due to adherent culture
Abstract translation: 本发明涉及用于从患者的肺癌组织中有效分离和增殖肺癌细胞的培养条件，以及使用由其培养的肺癌细胞筛选患者特异性癌症治疗剂的方法，更具体地，涉及 用于从肺癌组织分离和培养肺癌细胞的方法和使用通过该方法制备的肺癌细胞筛选患者特异性癌症治疗剂的方法。 从肺癌组织分离和培养肺癌细胞的方法包括以下步骤：1）将从患者分离的肺癌组织切成直径为40-60μm的大小; 2）允许步骤1）中的切割组织与待分离成单细胞的酶反应; 3）通过悬浮培养在低粘附培养皿中的无血清ACL4培养基或N 2添加培养基中的步骤2）中分离的单细胞增殖; 和4）使用Accutase，通过步骤3）中的悬浮培养物传代培养细胞，其中步骤4）不需要单独的Accutase反应终止步骤。 （附图标记）（A）根据现有技术从癌组织分离细胞; （AA）实体瘤; （B）根据本发明从肺癌组织中分离和培养肺癌细胞; （BB）胶原酶降解; （CC）单细胞悬液; （DD）琼脂中的细胞; （EE）孵育6-8天; （FF）细胞群体形成; （GG）克隆细胞分离; （HH）分离的克隆细胞重新分配; （二）切成40-100um的尺寸; （JJ）在超低粘附（或pluronic F127涂层）培养皿中的悬浮培养; （KK）有效细胞在无血清培养基中形成和培养; （LL）使用Accutase的悬浮培养细胞传代培养; （MM）贴壁培养; （NN）悬浮培养; （OO）EPCAM表达（％）; （PP）粘附培养; （QQ）暂停文化; （RR）通过悬浮培养培养和增殖以抑制由粘附培养引起的EMT现象
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Patent Agency Ranking
公开(公告)号：KR101972766B1
公开(公告)日：2019-04-29
申请号：KR1020120041753
申请日：2012-04-20
Applicant: (주)차바이오텍 , 차의과학대학교 산학협력단
IPC: C12N5/0735 , C12N5/074 , C12N15/86
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5.发明公开인간 다능성 줄기세포 단일세포의 계대배양 방법, 및 이를 이용한 형질전환 인간 다능성 줄기세포 제조 방법 审中-实审用于人单核细胞干细胞的再培养的方法及使用其制备转化的人肺细胞干细胞的方法
公开(公告)号：KR1020130118674A
公开(公告)日：2013-10-30
申请号：KR1020120041753
申请日：2012-04-20
Applicant: (주)차바이오텍 , 차의과학대학교 산학협력단
IPC: C12N5/074 , C12N5/0735 , C12N5/02 , C12N15/86
Abstract: PURPOSE: A subculture method of human pluripotent stem cells (hPSCs) is provided to allow isolated hPSCs, hart to culture, to form cell colonies and to be subcultured with maintaining pluripotency, so that the clonal isolated hPSCs are used for follow-up studies such as gene transduction, differentiation induction, etc. CONSTITUTION: A subculture method of hPSC single cells comprises: a step of preparing isolated hPSC single cells; and a step of enhancing colony formation potency by suppressing Gi-mediated GPCR signal transduction in the hPSC single cells. A manufacturing method of hPSC colonies comprises: a step of preparing isolated hPSC single cells; and a step of enhancing colony formation potency by suppressing Gi-mediated GPCR signal transduction in the hPSC single cells. [Reference numerals] (AA) Single hESC; (BB) Cell group forming area
Abstract translation: 目的：提供人多能干细胞（hPSC）的传代培养方法，以允许分离的hPSC，hart培养，形成细胞集落，并保持多能性进行传代培养，使得克隆分离的hPSC用于后续研究， 作为基因转导，分化诱导等。构成：hPSC单细胞的传代培养方法包括：制备分离的hPSC单细胞的步骤; 以及通过抑制hPSC单细胞中的Gi介导的GPCR信号转导来增强集落形成能力的步骤。 hPSC菌落的制备方法包括：制备分离的hPSC单细胞的步骤; 以及通过抑制hPSC单细胞中的Gi介导的GPCR信号转导来增强集落形成能力的步骤。 （附图标记）（AA）单个hESC; （BB）细胞组形成区
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公开(公告)号：KR101119464B1
公开(公告)日：2012-03-16
申请号：KR1020100078202
申请日：2010-08-13
Applicant: (주)차바이오텍 , 연세대학교 산학협력단
IPC: C12N5/0735 , C12N5/0775 , C12N5/02 , C07K14/475
Abstract: 본 발명은 인간 배아줄기세포를 결합 조직 성장 인자(connective tissue growth factor, CTGF) 및 소태아혈청(fetal bovine serum, FBS)을 포함하는 배지 중에서 배양하는 것을 포함하는, 인간 배아줄기세포의 중간엽 줄기세포로의 분화방법을 제공한다. 또한, 본 발명은 CTGF 및 FBS를 포함하는, 인간 배아줄기세포의 중간엽 줄기세포로의 분화용 배지를 제공한다. 또한, 본 발명은 인간 배아줄기세포의 중간엽 줄기세포로의 분화용 배지 및 중간엽 줄기세포의 배양용 배지를 제공한다.
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公开(公告)号：KR1020120021699A
公开(公告)日：2012-03-09
申请号：KR1020100078202
申请日：2010-08-13
Applicant: (주)차바이오텍 , 연세대학교 산학협력단
IPC: C12N5/0735 , C12N5/0775 , C12N5/02 , C07K14/475
Abstract: PURPOSE: A method for differentiating human embryonic stem cells into mesenchymal stem cells is provided to enhance differentiation efficiency. CONSTITUTION: A method for differentiating human embryonic stem cells into mesenchymal stem cells comprises a step of culturing the human embryonic stem cells in a medium containing 80-120 ng/ml of CTGF(connective tissue growth factor) and 2-5 w/v% of FBS(fetal bovine serum) for 2-5 days with daily fresh medium.
Abstract translation: 目的：提供一种将人胚胎干细胞分化为间充质干细胞的方法，以提高分化效率。 构成：将人胚胎干细胞分化为间充质干细胞的方法包括在含有80-120ng / ml CTGF（结缔组织生长因子）和2-5w / v％的培养基中培养人胚胎干细胞的步骤， 的FBS（胎牛血清）2-5天，每日新鲜培养基。
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