摘要
胚胎生殖（EG）细胞是来源于胚胎原始生殖细胞（PGCs）的多潜能干细胞。采用无饲养层细胞、无细胞因子等的基础培养液（DMEM＋20％NBS＋0．1mM 2Me）培养EG细胞，部分在基础培养液添加10μmol／LRA＋0．75％DMSO或10μmol／L 5-氮胞苷（5-AZA）诱导人类EG细胞向心肌细胞分化，以检测其是否具有向心肌细胞自发分化的能力。9例（8．49％，9／106）胎儿EG细胞在体外分化得到20个节律性心脏跳动样细胞团，其搏动节律为20～120次／min，体外维持节律性搏动最短2d，最长至15d，呈PAS，Myoglobin，α-actin阳性；对K^＋、Ca^＋、肾上腺素等具有与在体心脏相似的反应性；透射电镜观察具有心肌细胞样结构。添加DMSO和RA或5-AZA诱导未得到跳动样心肌细胞，但可提高心肌α-actin免疫组织化学染色阳性率；表明人类胚胎生殖细胞具有向心肌细胞分化的潜能。
Embryonic germ （EG） cells are pluripotent cells derived from primordial germ cells（PGCs） of gonads, gonadal ridges and mesenteries, analogies of fetuses,with the ability to undergo both highly self-renewal and multiple differentiation. These cells in vitro can differentiate into derivatives of all three embryonic germ layers when transferred to an in vitro environment and have the ability to form any fully differentiated cells of the body. The aim of this study is to investigate the potentiality of human EG cells differentiation into cardiomyocytes. Inducing human EG cells with the method of murine ES cells differentiation into cardiomyocytes,supplemented with 0.75%-1% DMSO,20%NES, 10^-7mM RA and 20% cardiomyocytes conditioned medium. Results：20 heartlike （rhythmic beating cell masses were observed in vitro culture and delayed human EG cells, which beat spontaneously from 20-120 times per minute and maintained beating for 2-15 days, periodic acid＇s staining （PAS）, Myogoblin and α-actin immunological histology positive were all positive and reacted with K^＋,Ca^2＋ and adrenalin. Relatively unorganized myofibrillar bundles or more organized sarcomeres, z-bands or a gap junction, the presence of desmosomes in a few cells of the cell masses was observed with transmision electron microscope ,which initially demonstrated that these cells were cardiomyocytes. We could not get rhythmly beating cardiomyocytes with 0.75%-1%DMSO,10^-7mM RA and 20% cardiomyocytes conditioned medium, but in which the percentage of cardiac α-actin immunostaining positive cells were increased. The results first demonstrated that human EG cells can differentiate into rhythmic beating cardiomyocytes in vitro and suggests that human EG cells may represent a new potent resource for cardiacomyocytes transplantation therapy for myocardium infarction.
出处
《生物医学工程学杂志》
EI
CAS
CSCD
北大核心
2006年第5期1080-1085,共6页
Journal of Biomedical Engineering
基金
国家自然科学基金资助项目(30200137)
国家高技术研究与发展计划(863)项目(2002AA216161
2005AA219050)
教育部重大科技项目(03160)
陕西省重大科技计划
西北农林科技大学青年学术骨干基金资助
关键词
原始生殖细胞
胚胎生殖细胞
胚胎干细胞
心肌细胞
Primordial germ cells（PGCs） Embryonic germ（EG） cells Embryonic stem（ES） cells Cardiomyocytes
