Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

Clonal derivation and characterization of human embryonic stem cell lines

Heins, N ; Lindahl, A ; Karlsson, U ; Rehnstrom, M ; Caisander, G ; Emanuelsson, K ; Hanson, C ; Semb, Henrik LU ; Bjorquist, P and Sartipy, P , et al. (2006) In Journal of Biotechnology 122(4). p.511-520
Abstract
Human embryonic stem cells (hESC) are isolated as clusters of cells from the inner cell mass of blastocysts and thus should formally be considered as heterogeneous cell populations. Homogenous hESC cultures can be obtained through subcloning. Here, we report the clonal derivation and characterization of two new hESC lines from the parental cell line SA002 and the previously clonally derived cell line AS034.1, respectively. The hESC line SA002 was recently reported to have an abnormal karyotype (trisomy 13), but within this population of cells we observed rare individual cells with an apparent normal karyotype. At a cloning efficiency of 5%, we established 33 subclones from SA002, out of which one had a diploid karyotype and this subline... (More)
Human embryonic stem cells (hESC) are isolated as clusters of cells from the inner cell mass of blastocysts and thus should formally be considered as heterogeneous cell populations. Homogenous hESC cultures can be obtained through subcloning. Here, we report the clonal derivation and characterization of two new hESC lines from the parental cell line SA002 and the previously clonally derived cell line AS034.1, respectively. The hESC line SA002 was recently reported to have an abnormal karyotype (trisomy 13), but within this population of cells we observed rare individual cells with an apparent normal karyotype. At a cloning efficiency of 5%, we established 33 subclones from SA002, out of which one had a diploid karyotype and this subline was designated SA002.5. From AS034.1 we established one reclone designated AS034. 1.1 at a cloning efficiency of 0.1%. These two novel sublines express cell surface markers indicative of undifferentiated hESC (SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81), Oct-4, alkaline phosphatase, and they display high telomerase activity. In addition, the cells are pluripotent and form derivatives of all three embryonic germ layers in vitro as well as in vivo. These results, together with the clonal character of SA002.5 and AS034. 1.1 make these homogenous cell populations very useful for hESC based applications in drug development and toxicity testing. In addition, the combination of the parental trisomic hESC line SA002 and the diploid subclone SA002.5 provides a unique experimental system to study the molecular mechanisms underlying the pathologies associated with trisomy 13. (c) 2005 Elsevier B.V. All rights reserved. (Less)
Please use this url to cite or link to this publication:
author
; ; ; ; ; ; ; ; and , et al. (More)
; ; ; ; ; ; ; ; ; and (Less)
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
characterization, human embryonic stem cells, subcloning
in
Journal of Biotechnology
volume
122
issue
4
pages
511 - 520
publisher
Elsevier
external identifiers
  • pmid:16324761
  • wos:000236823800011
  • scopus:33645378510
  • pmid:16324761
ISSN
1873-4863
DOI
10.1016/j.jbiotec.2005.10.010
language
English
LU publication?
yes
additional info
The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Stem Cell and Pancreas Developmental Biology (013212044)
id
196ae5b5-f15e-4ce0-aad5-7b42ae1681d4 (old id 414032)
date added to LUP
2016-04-01 12:24:03
date last changed
2021-03-03 05:14:04
@article{196ae5b5-f15e-4ce0-aad5-7b42ae1681d4,
  abstract     = {Human embryonic stem cells (hESC) are isolated as clusters of cells from the inner cell mass of blastocysts and thus should formally be considered as heterogeneous cell populations. Homogenous hESC cultures can be obtained through subcloning. Here, we report the clonal derivation and characterization of two new hESC lines from the parental cell line SA002 and the previously clonally derived cell line AS034.1, respectively. The hESC line SA002 was recently reported to have an abnormal karyotype (trisomy 13), but within this population of cells we observed rare individual cells with an apparent normal karyotype. At a cloning efficiency of 5%, we established 33 subclones from SA002, out of which one had a diploid karyotype and this subline was designated SA002.5. From AS034.1 we established one reclone designated AS034. 1.1 at a cloning efficiency of 0.1%. These two novel sublines express cell surface markers indicative of undifferentiated hESC (SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81), Oct-4, alkaline phosphatase, and they display high telomerase activity. In addition, the cells are pluripotent and form derivatives of all three embryonic germ layers in vitro as well as in vivo. These results, together with the clonal character of SA002.5 and AS034. 1.1 make these homogenous cell populations very useful for hESC based applications in drug development and toxicity testing. In addition, the combination of the parental trisomic hESC line SA002 and the diploid subclone SA002.5 provides a unique experimental system to study the molecular mechanisms underlying the pathologies associated with trisomy 13. (c) 2005 Elsevier B.V. All rights reserved.},
  author       = {Heins, N and Lindahl, A and Karlsson, U and Rehnstrom, M and Caisander, G and Emanuelsson, K and Hanson, C and Semb, Henrik and Bjorquist, P and Sartipy, P and Hyllner, J},
  issn         = {1873-4863},
  language     = {eng},
  number       = {4},
  pages        = {511--520},
  publisher    = {Elsevier},
  series       = {Journal of Biotechnology},
  title        = {Clonal derivation and characterization of human embryonic stem cell lines},
  url          = {http://dx.doi.org/10.1016/j.jbiotec.2005.10.010},
  doi          = {10.1016/j.jbiotec.2005.10.010},
  volume       = {122},
  year         = {2006},
}