Scientists with the International Stem Cell Initiative (ISCI) have identified a set of common gene expression markers that may be used to characterise diverse human embryonic stem cell (hESC) lines.
The initiative represents the first internationally coordinated effort to provide a systematic and comparative survey of the hESC lines available from various research groups around the world. The results of the study were published in the July issue of Nature Biotechnology.
ISCI employed technology from Applied Biosystems to carry out the first comparative study of a large and diverse collection of hESC lines from 17 laboratories in the U.S., Europe, Israel, Japan, and Australia. The team used several approaches to identify a reliable set of genetic markers to characterize hESCs, which included profiling 93 genes in 59 different cell lines.
A major finding from this first phase of the ICSI project was that all of the cell lines studied shared a number of key molecular signatures, despite different genetic constitutions and the various culture techniques to which they have been subjected. It is expected that these molecular markers may be used to validate the potential of individual hESC lines to differentiate into other types of cells, also referred to as their pluripotency status.
The ISCI scientists used the Applied Biosystems 7900HT Fast Real-Time PCR System and custom Applied Biosystems TaqMan® Arrays to identify six key genetic markers and 14 additional genes highly correlated with a key pluripotency marker in all the cell lines tested. Applied Biosystems worked with the ISCI researchers at the centralised analytical laboratory at the University of Sheffield in the U.K., to select the assays, process the arrays and analyse the results.
“These findings are a crucial first step in ensuring that future advances in the field of stem cell research involve internationally coordinated quality standards,” said Paul Gokhale, Ph.D, of the Centre for Stem Cell Research at the University of Sheffield. “Our collective aim is to provide openness, reliability, and the ability for scientists to confidently reproduce and extend each other’s work.”
As a result of this ISCI study, Applied Biosystems has developed the TaqMan Array Human Stem Cell Pluripotency Panel, which targets 96 genes for independent monitoring of hESC lines to help investigators compare cell lines derived from diverse sources.
The company said it has also created a TaqMan Array Mouse Stem Cell Pluripotency Panel comprised of mouse orthologs to the genes on the human panel.