Characterization and gene expression profiling of five new human embryonic stem cell lines derived in Taiwan
Characterization and gene expression profiling of five new human embryonic stem cell lines derived in Taiwan
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DOI：
10.1089/scd.2006.15.532
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发表时间：
2006-08-01
影响因子：
4
通讯作者：
Singh, Sher
中科院分区：
文献类型：
--
作者：
Li, Steven Shoei-Lung;Liu, Yung-Hsien;Singh, Sher
许多人胚胎干细胞（hESC）系已被报道，但只有少数人已被充分表征。本研究从台湾32个废弃胚泡中获得5个新的hESC系，这些系在hESC培养基中的有丝分裂失活的小鼠胚胎成纤维细胞（MEF）饲养层上连续培养超过44代，并经历冻融过程。所有五个hESC系均表达特征性未分化hESC标记物，如SSEA-4、TRA-1-81、碱性磷酸酶、TERT以及转录因子POU 5 F1（OCT 4）和NANOG。人胚胎干细胞T1和T3系具有正常的雌性核型，而T4和T5系是正常的雄性核型，而T2系是雄性12三体（47 XY，+12）。人胚胎干细胞T1、T2、T3和T5系能在严重联合免疫缺陷（SCID）小鼠体内产生畸胎瘤，T4系只能在体外形成胚状体。使用Affytelium人类基因组U133 plus 2.0 GeneChip分析这五个新衍生的hESC系的全局基因表达谱。结果显示，在所有5个hESC系中检测到4，145个转录本，包括19%的未知功能。将5个hESC系中普遍表达的4，145个基因与hESC系T1和胎盘产生的畸胎瘤中表达的那些基因进行比较，发现40个基因在所有5个hESC系中特异表达。这40个基因包括先前报道的干细胞基因，如POU 5 F1（OCT 4），NANOG，TDGF 1（CRIPTO），SALL 4，LECT 1和BUB 1负责自我更新和多能分化。整体基因表达分析还表明，转化生长因子-β（TGF-β）/激活素分支组分TGF-BC、ACVR 2A、ACVR 1（ALK 2）、TGF-β R 1（ALK 5）和SMAD 2在这五种hESC系的未分化状态中高度表达，并在分化后降低。简而言之，这五种hESC系的hESC性质得到未分化状态、广泛的更新能力和多能性（包括形成畸胎瘤和/或EB的能力）的支持。这些细胞系将用于人类胚胎干细胞生物学和药物开发。
Many human embryonic stem cell (hESC) lines have been reported, but only a few of them have been fully characterized. In this report, five new hESC lines were derived from 32 discarded blastocysts in Taiwan, and these lines were continuously cultured on mitotically inactivated mouse embryonic fibroblast (MEF) feeder layer in the hESC medium for more than 44 passages and underwent freezing/thawing processes. All five hESC lines expressed characteristic undifferentiated hESC markers, such as SSEA-4, TRA-1-81, alkaline phosphatase, TERT, and the transcription factors POU5F1 (OCT4) and NANOG. hESC lines T1 and T3 possess normal female karyotypes, whereas lines T4 and T5 are normal male, but line T2 is male trisomy 12 (47XY, +12). hESC lines T1, T2, T3, and T5 were able to produce teratomas in severe combined immunodeficient (SCID) mice, and line T4 could only form embryoid bodies (EBs) in vitro. Global gene expression profiles of these five newly derived hESC lines were analyzed using the Affymetrix human genome U133 plus 2.0 GeneChip. The results showed that 4,145 transcripts, including 19% of unknown functions, were detected in all five hESC lines. Comparison of the 4,145 genes commonly expressed in the five hESC lines with those genes expressed in teratomas produced by the hESC line T1 and placenta revealed 40 genes exclusively expressed in all five hESC lines. These 40 genes include the previously reported stemness genes, such as POU5F1 (OCT4), NANOG, TDGF1 (CRIPTO), SALL4, LECT1, and BUB1 responsible for self-renewal and pluripotent differentiation. The global gene expression analysis also indicated that the transforming growth factor-beta (TGF-beta)/activin branch components inhibin BC, ACVR2A, ACVR1 (ALK2), TGFBR1 (ALK5), and SMAD2 were found to be highly expressed in undifferentiated states of these five hESC lines and decreased upon differentiation. In short, the hESC nature of these five hESC lines is supported by the undifferentiated state, extensive renewal capacity, and pluripotency, including the ability to form teratomas and/or EBs. These cell lines will be useful for human embryonic stem cell biology and drug development.