A molecular scheme for improved characterization of human embryonic stem cell lines
1 Stem Cell Center, American Type Culture Collection (ATCC®), Manassas, VA, USA
2 Cell Biology Department, American Type Culture Collection (ATCC®), Manassas, VA, USA
3 Laboratory of Neuroscience, National Institute on Aging, National Institutes of Health, Baltimore, MD, USA
4 Buck Institute for Age Research, Novato, CA, USA
5 Burnham Institute, La Jolla, CA, USA
6 Departments of Pathology, Oncology, and Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA
7 Invitrogen Corporation, Carlsbad, CA, USA
BMC Biology 2006, 4:28 doi:10.1186/1741-7007-4-28Published: 18 August 2006
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.
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.
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.