公开(公告)号：KR100856706B1

公开(公告)日：2008-09-04

申请号：KR1020060110269

申请日：2006-11-09

Applicant: 고려대학교 산학협력단

Inventor： 김종훈 ,  김병각

IPC: C12N5/071 ,  C12N5/07 ,  C12N5/02

Abstract: 본 발명은 인간배아 줄기세포로부터 형성된 배아체를 혈관내피 성장인자 (VEGF)로 처리하여 신경외배엽이 증가된 배아체로 유도함으로써 신경세포로의 분화를 촉진시키는 방법에 관한 것이다.
본 발명에 따르면, 인간배아 줄기세포를 혈관내피 성장인자를 이용하여 신경외배엽이 증가된 배아체로 유도함으로써, 중배엽, 내배엽으로부터 유래되는 심근세포, 근육세포, 간세포, 조혈세포 등 각종의 세포로 분화할 가능성을 원천적으로 봉쇄하여 세포치료를 위한 순도 높은 신경세포군으로의 분화효율을 증가시킬 수 있다.
줄기세포, 배아체, 혈관내피 성장인자, 신경세포

公开(公告)号：KR1020080023078A

公开(公告)日：2008-03-12

申请号：KR1020060110269

申请日：2006-11-09

Applicant: 고려대학교 산학협력단

Inventor： 김종훈 ,  김병각

IPC: C12N5/071 ,  C12N5/07 ,  C12N5/02

Abstract: Methods for inducing differentiation from human embryonic stem cells to dopaminergic neurons are provided to block possibility of differentiation into various cells including myocardial cells, muscle cells, liver cells and hematopoietic cells, so that differentiation efficiency into high purity nerve cells for cell therapy is increased. A method for inducing differentiation from human embryonic stem cells to dopaminergic neurons comprises the steps of: treating embryoid bodies formed from human embryonic stem cells with 40-60 ng/ml of a vascular endothelial growth factor(VEGF) for 7-9 days to induce the neuroectoderm-increased embryoid bodies; culturing the neuroectoderm-increased embryoid bodies attached to a tissue culture dish to induce neural precursor cells; detaching the induced neural precursor cells from the tissue culture dish and re-attaching and culturing them in the dish to increase purity and maintain density uniformly; and finally differentiating the high purity of neural precursor cells to dopaminergic neurons. Further, a 20-30 ug/ml of fibronectin is added into the neuroectoderm-increased embryoid bodies.

Abstract translation: 提供诱导从人胚胎干细胞分化为多巴胺能神经元的方法，以阻断分化成包括心肌细胞，肌细胞，肝细胞和造血细胞在内的各种细胞的可能性，从而增加细胞治疗的高纯度神经细胞的分化效率。 诱导从人胚胎干细胞分化为多巴胺能神经元的方法包括以下步骤：用40-60ng / ml血管内皮生长因子（VEGF）处理由人胚胎干细胞形成的胚状体7-9天以诱导 神经外胚层增生的胚状体; 培养附着于组织培养皿的神经外胚层增加的胚状体以诱导神经前体细胞; 从组织培养皿中分离诱导的神经前体细胞，并将其重新附着并培养在培养皿中以提高纯度并均匀地保持密度; 最后将神经前体细胞的高纯度区分为多巴胺能神经元。 此外，将20-30μg/ ml的纤连蛋白加入到神经外胚层增加的胚状体中。