公开(公告)号：KR102224273B1

公开(公告)日：2021-03-08

申请号：KR1020190125458

申请日：2019-10-10

Applicant: 고려대학교 산학협력단

Inventor： 임도선 ,  최승철 ,  주형준 ,  김종호 ,  박치연 ,  박용두

IPC: C12N5/077 ,  G01N33/50

Abstract: 본발명은줄기세포유래성숙심근세포및 이를이용한심혈관질환모델에관한것으로, 더욱자세하게는 FGF4 및아스코르브산을함유하는배지에서줄기세포를배양하여성숙심실심근세포로분화시키고, 분화된성숙심실심근세포를심혈관질환세포모델로활용하는것에관한것이다. 본발명에따른 FGF4 및아스코르브산을함유하는배지에서줄기세포를배양하여분화유도한성숙심실심근세포및 이를이용한심혈관질환세포모델은심혈관질환치료제스크리닝및 신약독성평가에매우유용하다.

公开(公告)号：KR101881531B1

公开(公告)日：2018-07-25

申请号：KR1020160158414

申请日：2016-11-25

Applicant: 고려대학교 산학협력단

Inventor： 임도선 ,  김종호 ,  주형준

IPC: C12N5/0775 ,  C12P21/00 ,  C07K14/52

Abstract: 본발명은폴리-2-하이드록시에틸메타아크릴레이트 (Poly-2-hydroxyethyl methacrylate; Poly-HEMA)를이용하여사이토카인또는성장인자의발현능을증가시킨중간엽줄기세포의제조방법에관한것이다. 본발명에따른폴리-2-하이드록시에틸메타아크릴레이트를이용하여사이토카인또는성장인자의발현능을증가시킨중간엽줄기세포의제조방법은생체이외의성분을이용하지않고줄기세포를분화시킬수 있으므로, 임상적으로부작용없는줄기세포치료에매우유용하다.

公开(公告)号：KR1020180022119A

公开(公告)日：2018-03-06

申请号：KR1020160107023

申请日：2016-08-23

Applicant: 고려대학교 산학협력단

Inventor： 임도선 ,  이규백 ,  주형준 ,  서하림 ,  최룡휘

IPC: C12N5/077 ,  C12N5/00

CPC classification number: C12N5/00 ,  C12N5/0657 ,  C12N5/0068 ,  C12N2501/19 ,  C12N2501/998 ,  C12N2506/45 ,  C12N2535/00

Abstract: 본발명은줄기세포를나노패턴배양접시에배양하여심근세포로분화시키는방법에관한것으로, 더욱자세하게는직경이일정하게증가하고간격이일정하게감소하는이중구배나노패턴배양접시를이용하여줄기세포를심근세포로분화시키는방법에관한것이다. 본발명에따르면, 심근세포분화에최적화된나노패턴배양접시를이용한줄기세포를심근세포로분화시키는방법은고가의세포외기질을코팅하거나지지세포를이용할필요없어경제적이며, 심근세포로의분화효율이우수하다.

Abstract translation: 本发明涉及一种用于干细胞分化成心肌细胞的方法通过培养的纳米图案培养板，并且更特别地，使用这种介质倍纳米图案培养皿心脏干细胞，以减少在yiiljeong直径和间距yiiljeong的增加 到一个单元格。 根据本发明，使用用于心肌细胞进行了优化的板线干细胞分化的纳米图案培养物的方法分化成心肌细胞，且经济，无需涂层或使用饲养细胞的心肌细胞的细胞外基质昂贵细胞分化效率 赢得。

公开(公告)号：KR100973823B1

公开(公告)日：2010-08-04

申请号：KR1020080008364

申请日：2008-01-28

Applicant: 고려대학교 산학협력단

Inventor： 최승철 ,  최지현 ,  임도선

IPC: C12N5/071 ,  C12N5/02 ,  C12N5/00

Abstract: 본 발명은 배아성 암종 세포를 기본배지 및 혈청을 포함하나 되먹임세포(feeder cell) 및 분화억제제가 첨가되지 않은 배양배지에서 배양하여 배상체를 형성하는 단계 및 상기 형성된 배상체에 혈관내피성장인자(Vascular Endothelial Growth Factor)를 처리하여 혈관내피세포로 분화 유도하는 단계를 포함하는 배아성 암종 세포로부터 혈관내피세포로의 분화유도방법에 관한 것이다.
본 발명에 따르면, 배아성 암종 세포가 혈관내피세포로 분화능이 있음을 혈관내피세포 마커를 사용한 면역염색법, 실시간 중합효소연쇄반응(real-time PCR)에 의한 혈관내피세포 마커들의 정량적 분석, 배양액 조성 및 우태아혈청 첨가 농도에 따른 혈관내피세포로의 분화율 비교, 유세포 분석을 통하여 정량적으로 입증하였으며, 이러한 결과를 토대로 배아성 암종 세포를 혈관내피세포로 분화시켜 혈관내피세포를 대량으로 배양할 수 있다. 배아성 암종 세포는 유전자 도입이 쉽고, 세포 배양이 용이하며, 저비용으로 대량 배양이 가능한 장점을 가지고 있기 때문에 본 발명에 따라 생산된 혈관세포를 이용하여 혈관내피세포로의 분화 기전 연구, 심혈관 및 뇌혈관질환의 치료 연구, 신생혈관 차단에 의한 항암제 개발 연구 등에 유용하게 이용할 수 있다.

公开(公告)号：KR1020090055964A

公开(公告)日：2009-06-03

申请号：KR1020070122869

申请日：2007-11-29

Applicant: 고려대학교 산학협력단

Inventor： 임도선 ,  최승철

IPC: C12N5/07

Abstract: A method for isolating a mesenchymal stem cell which is derived from a bone marrow is provided to isolate the mesenchymal stem cell of high purity using an isolation marker, Sca-1, and improve the cell therapy by differentiating to a cardiac myocyte, adipocyte, and osteocyte. A method for isolating a mesenchymal stem cell derived from a bone marrow comprises: a step of isolating a myelocyte from the bone marrow and attaching-culturing in a culture container; a step of treating a trypsin/ethylenediaminetetraacetic acid(EDTA) solution into the myelocyte and attaching-culturing a myelocyte isolated by the trypsin/EDTA solution in another culture container; a step of reacting with a linker-conjugated Sca-1 antibody; a step of treating with an anti-linker magnetic bead; and a step of separating the myelocyte which is bonded with the anti-linker magnetic bead using a magnet-activated column.

Abstract translation: 提供了分离源自骨髓的间充质干细胞的方法，以使用分离标记Sca-1分离高纯度的间充质干细胞，并通过区分心肌细胞，脂肪细胞和 骨细胞。 用于分离源自骨髓的间充质干细胞的方法包括：从骨髓中分离骨髓细胞并在培养容器中附着培养的步骤; 将胰蛋白酶/乙二胺四乙酸（EDTA）溶液处理到骨髓细胞中并将由胰蛋白酶/ EDTA溶液分离的骨髓细胞附着培养在另一培养容器中的步骤; 与接头偶联的Sca-1抗体反应的步骤; 用反接头磁珠处理的步骤; 以及使用磁激活柱分离与抗连接磁珠结合的骨髓细胞的步骤。

公开(公告)号：KR1020090055953A

公开(公告)日：2009-06-03

申请号：KR1020070122850

申请日：2007-11-29

Applicant: 고려대학교 산학협력단

Inventor： 임도선 ,  최승철

IPC: C12N15/64 ,  C12N15/09 ,  C12Q1/68

CPC classification number: C12N15/63 ,  C12Q1/6816 ,  C12Q1/6897

Abstract: A recombinant expression vector for monitoring the differentiation of stem cell to a cardiomyogenic cell is provided to detect the differentiation and use for cell therapy of cardiovascular disease and stability and efficiency of stem cell therapy. A recombinant expression vector pDsRed-Express-1/mMLC, pDsRed-Express-1/rMLC, pEGFP-1/cTn-1, or pDsRed-Express-1/cTn-1 for monitoring the differentiation to a cardiomyogenic cell comprises a promoter inducing the cardiomyogenic cell-specific expression and reporter gene which is operably linked. The promoter inducing the cardiomyogenic cell-specific expression is the promoter myosin light chain 2v promoter of the sequence number 1(SEQ ID NO:1), rat myosin light chain 2v promoter of the sequence number 2, or myocardial troponin I promoter of the sequence number 3. A method for monitoring the differentiation to cardiomyogenic cell comprises: a step of preparing a mesenchymal stem cell; a step of transducing the recombinant expression vector into a prepared stem cell; and a step of confirming the expression of reporter gene in the transduced stem cell.

Abstract translation: 提供了一种用于监测干细胞向致心肌细胞分化的重组表达载体，用于检测心血管疾病细胞治疗的分化和用途以及干细胞治疗的稳定性和效率。 用于监测分化成心肌细胞的重组表达载体pDsRed-Express-1 / mMLC，pDsRed-Express-1 / rMLC，pEGFP-1 / cTn-1或pDsRed-Express-1 / cTn-1包含启动子诱导 心肌细胞特异性表达和可操作地连接的报告基因。 诱导心肌细胞特异性表达的启动子是序列号1（SEQ ID NO：1），序列号2的大鼠肌球蛋白轻链2v启动子或序列的心肌肌钙蛋白I启动子的启动子肌球蛋白轻链2v启动子 3.一种监测分化成心肌细胞的方法，包括：制备间充质干细胞的步骤; 将重组表达载体转导到制备的干细胞中的步骤; 以及确认转录干细胞中报道基因表达的步骤。

公开(公告)号：KR1020090055869A

公开(公告)日：2009-06-03

申请号：KR1020070122718

申请日：2007-11-29

Applicant: 고려대학교 산학협력단

Inventor： 최승철 ,  임도선

IPC: C12N15/64 ,  C12N15/09 ,  C12Q1/68

CPC classification number: C12N15/63 ,  C12Q1/6816 ,  C12Q1/6897

Abstract: A recombinant expression vector for monitoring the differentiation of a cardiomyogenic cell and endothelial cell is provided to detect the cell differentiation after cell therapy and verify the stability. A recombinant expression vector pMLC-2v-DsRed-Flk-1-EGFP or pMLC-2v-DsRed-Tie-2-EGFP for monitoring the differentiation of a cardiomyogenic cell and endothelial cell comprises a promoter which induces the cardiomyogenic cell-specific expression; a first reporter gene which is connected to be operatable; a promoter which induces the endothelial cell-specific expression; and a second reporter gene which is connected to be operatable. The promoter which induces cardiomyogenic cell-specific expression is the myosin light chain 2v promoter of the sequence number 1(SEQ ID NO:1), rat myosin light chain 2v promoter of the sequence number 2, or myocardium troponin I promoter of the sequence number 3. The endothelial cell-specific promoter is the Flk-1 promoter of the sequence number 4 and Tie-2 promoter of the sequence number 5. A method for monitoring the differentiation of cardiomyogenic cell and endothelial cell at once: a step of preparing a mesenchymal stem cell; a step of inducing the recombinant expression vector for monitoring the differentiation of cardiomyogenic cell and endothelial cell into the mesenchymal stem cell; and a step of confirming the expression of reporter gene in a transformed mesenchymal stem cell.

Abstract translation: 提供用于监测心肌细胞和内皮细胞分化的重组表达载体以检测细胞治疗后的细胞分化并验证稳定性。 用于监测心肌发生细胞和内皮细胞分化的重组表达载体pMLC-2v-DsRed-Flk-1-EGFP或pMLC-2v-DsRed-Tie-2-EGFP包含诱导心肌细胞特异性表达的启动子; 连接到可操作的第一报告基因; 诱导内皮细胞特异性表达的启动子; 和连接成可操作的第二报道基因。 引起心肌发生细胞特异性表达的启动子是序列号1（SEQ ID NO：1）的肌球蛋白轻链2v启动子，序列号2的大鼠肌球蛋白轻链2v启动子或序列号的心肌肌钙蛋白I启动子 3.内皮细胞特异性启动子是序列号为4的Flk-1启动子和序列号5的Tie-2启动子。一种用于一次性监测心肌细胞和内皮细胞分化的方法： 间充质干细胞 诱导重组表达载体监测心肌细胞和内皮细胞分化为间充质干细胞的步骤; 以及确认报道基因在转化的间充质干细胞中的表达的步骤。