A Modified Culture Medium Increases Blastocyst Formation and the Efficiency of Human Embryonic Stem Cell Derivation from Poor-Quality Embryos
A Modified Culture Medium Increases Blastocyst Formation and the Efficiency of Human Embryonic Stem Cell Derivation from Poor-Quality Embryos
复制标题
改良培养基可增加囊胚形成和从劣质胚胎衍生人胚胎干细胞的效率
DOI：
10.1262/jrd.09-225m
复制
发表时间：
2010-10-01
影响因子：
1.8
通讯作者：
Sun, Xiaofang
中科院分区：
文献类型：
--
作者：
Fan, Yong;Luo, Yumei;Sun, Xiaofang
人类胚胎干细胞（HESCs）被定义为自我更新的细胞，保留其分化为身体所有细胞类型的能力。它们在医学应用和作为早期人类发展的模型方面具有巨大的潜力。有必要衍生新的HESC系，以满足其在细胞替代疗法、疾病建模和基础研究中的应用需求。在这里，我们描述了一种含有人重组白血病抑制因子和人碱性成纤维细胞生长因子的改良培养基，该培养基显著提高了人囊胚的形成数量和质量，以及劣质胚胎HESC衍生的效率。在改良培养基中培养质量差的胚胎，囊胚形成率比在常规培养基中提高2倍，衍生效率提高7倍。我们从改良培养基中培养的劣质胚胎中获得了15个HESC系，从常规培养基中培养的优质胚胎中获得了2个HESC系。所有细胞系都具有典型的人类多能干细胞特征，包括相似的形态、正常的核型、碱性磷酸酶、多能基因（如Oct4）和细胞表面标记（SSEA-4、TRA-1-60、TRA-1-81）的表达、在SCID小鼠中形成畸胎瘤的能力，以及在体外分化为三种胚胎胚层细胞的能力。我们的数据表明，在我们改良的培养基中达到囊胚阶段的低质量胚胎是正常HESC系衍生的可靠来源。
Human embryonic stem cells (HESCs) are defined as self-renewing cells that retain their ability to differentiate into all cell types of the body. They have enormous potential in medical applications and as a model for early human development. There is a need for derivation of new HESC lines to meet emerging requirements for their use in cell replacement therapies, disease modeling, and basic research. Here, we describe a modified culture medium containing human recombinant leukemia inhibitory factor and human basic fibroblast growth factor that significantly increases the number of human blastocysts formed and their quality, as well as the efficiency of HESC derivation from poor-quality embryos. Culturing poor-quality embryos in modified medium resulted in a two-fold increase in the blastocyst formation rate and a seven-fold increase over the derivation efficiency in conventional medium. We derived 15 HESC lines from poor-quality embryos cultured in modified culture medium and two HESC lines from quality embryos cultured in conventional culture medium. All cell lines shared typical human pluripotent stem cell features including similar morphology, normal karyotypes, expression of alkaline phosphatase, pluripotency genes, such as Oct4, and cell surface markers (SSEA-4, TRA-1-60, TRA-1-81), the ability to form teratomas in SCID mice, and the ability to differentiate into cells of three embryonic germ layers in vitro. Our data suggest that poor-quality embryos that have reached the blastocyst stage in our modified culture medium are a robust source for normal HESC line derivation.