A molecular scheme for improved characterization of human embryonic stem cell lines.
BACKGROUND
Human embryonic stem cells (hESC) offer a renewable source of a wide range of cell types for use in research and cell-based therapies to treat disease. Inspection of protein markers provides important information about the current state of the cells and data for subsequent manipulations. However, hESC must be routinely analyzed at the genomic level to guard against deleterious changes during extensive propagation, expansion, and manipulation in vitro.


RESULTS
We found that short tandem repeat (STR) analysis, human leukocyte antigen (HLA) typing, single nucleotide polymorphism (SNP) genomic analysis, mitochondrial DNA sequencing, and gene expression analysis by microarray can be used to fully describe any hESC culture in terms of its identity, stability, and undifferentiated state.


CONCLUSIONS
Here we describe, using molecular biology alone, a comprehensive characterization of 17 different hESC lines. The use of amplified nucleic acids means that for the first time full characterization of hESC lines can be performed with little time investment and a minimum of material. The information thus gained will facilitate comparison of lines and replication of results between laboratories.
Josephson
Richard
R
Stem Cell Center, American Type Culture Collection (ATCC), Manassas, VA, USA. rjosephson@globalstem.com
Sykes
Gregory
G
Liu
Ying
Y
Ording
Carol
C
Xu
Weining
W
Zeng
Xianmin
X
Shin
Soojung
S
Loring
Jeanne
J
Maitra
Anirban
A
Rao
Mahendra S
MS
Auerbach
Jonathan M
JM
D002454:Cell Differentiation; D002460:Cell Line; D018920:Coculture Techniques; D053595:Embryonic Stem Cells / Q000166:cytology*; D005819:Genetic Markers; D006680:HLA Antigens / Q000235:genetics; D006801:Humans; D018895:Microsatellite Repeats; D020411:Oligonucleotide Array Sequence Analysis; D039904:Pluripotent Stem Cells / Q000166:cytology*; D020641:Polymorphism, Single Nucleotide
D005819:Genetic Markers; D006680:HLA Antigens