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Human embryonic stem cells (hESCs) possess enormous capability serving as an excellent model for human embryonic development, as a unique platform for pharmaceutical screening and as an

unlimited resource for cell replacement therapy. Currently, worldwide attention has focused on the derivation of hESCs for clinical application. However, the majority of already established

hESCs including NIH-approved lines have been directly or indirectly exposed to non-human materials during their derivation and/or propagation, which greatly restrict their future therapeutic

potential. Although the human-feeder systems, the autogeneic-feeder systems and multiple feeder-free systems may be safe for cultivation of undifferentiated hESCs, the derivation procedure

free of xeno-products needs to be developed. Here we adopted a non-contact laser-assisted hatching system in combination with sequential culture process to obtain hatched blastocysts as

materials for hESC derivation, and derived a well-characterized hESC line, ZJUhES-1, from a laser-assistedly hatched blastocyst of Chinese population without exposure to any non-human

materials during derivation. The cell line satisfies the criteria of pluripotent hESCs: typically morphological characteristics; the expression of alkaline phosphatase, human telomerase

reverse transcriptase and a series of hESC-specific markers including SSEA-3, SSEA-4, TRA-1-60, TRA-1-81, OCT-4, NANOG, REX-1, SOX-2, UTF-1, CONNEXINS 43 and 45, TERF-1 and TERF-2, GLUT-1,

BCRP-1/ ABCG-2, GDF3, LIN28, FGF4, Thy1, Cripto1/TDGF1, AC133 as well as SMAD2/3 and SMAD1/5 which are the signal transducers of TGFa superfamily signaling pathway that is required for

maintaining hESC identity; extended proliferative capacity; maintenance of a stable male karyotype after long-term cultivation; robust multiple-lineage developmental potentials to form

derivatives of all three embryonic germ layers both _in vivo_ and _in vitro_. Moreover, the cell line has distinct identity revealed from DNA fingerprinting. In addition, accumulating

evidences suggest that directed differentiation of hESCs toward a homogenous population of a specific cell type is prerequisite for their unlimited application in regenerative medicine.

However, most published differentiation protocols involving embryoid body formation and the use of coculture stromal cells are poorly defined and inefficient. Starting from monolayer

culture, we use all-trans retinoic acid plus FGFs and different culture media to allow for the selective induction of hESCs toward neural, glia and early smooth muscle lineages,

demonstrating the pluripotency of ZJUhES-1 in an easy manipulation way. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * College of Life Science, Zhejiang University, Hangzhou, 310058, China

Rongrong Wu, Xiaoli Zhao & Ming Zhang * In Vitro Fertilization Center of Women's Hospital, College of Medicine, Zhejiang University, Hangzhou, 310058, China Chenming Xu & Fan

Jin * Institute of Genetics and Developmental Biology, Chinese Academy of Science, Beijing, 100101, China Liangbiao Chen * Central Hospital of Huzhou, Huzhou, 313000, China Xing Yao & 

Licheng Dai Authors * Rongrong Wu View author publications You can also search for this author inPubMed Google Scholar * Chenming Xu View author publications You can also search for this

author inPubMed Google Scholar * Liangbiao Chen View author publications You can also search for this author inPubMed Google Scholar * Xiaoli Zhao View author publications You can also

search for this author inPubMed Google Scholar * Xing Yao View author publications You can also search for this author inPubMed Google Scholar * Licheng Dai View author publications You can

also search for this author inPubMed Google Scholar * Fan Jin View author publications You can also search for this author inPubMed Google Scholar * Ming Zhang View author publications You

can also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to Ming Zhang. RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS

ARTICLE Wu, R., Xu, C., Chen, L. _et al._ Derivation, characterization and directed differentiation of a new human embryonic stem cell line from a Chinese blastocyst hatched by a non-contact

laser system. _Cell Res_ 18 (Suppl 1), S134 (2008). https://doi.org/10.1038/cr.2008.224 Download citation * Published: 04 August 2008 * Issue Date: August 2008 * DOI:

https://doi.org/10.1038/cr.2008.224 SHARE THIS ARTICLE Anyone you share the following link with will be able to read this content: Get shareable link Sorry, a shareable link is not currently

available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing initiative KEYWORDS * human embryonic stem cells * derivation * hatched blastocysts *

laser-assisted hatching system * non-contact laser * directed differentiation