Fluidigm Europe's paper 'Single-Embryo Gene Expression for Early Embryo Development' describes the work of Dr Mylene Yao and her fellow researchers at Stanford University.
Many scientists are currently attempting to reprogram a highly differentiated somatic cell into a pluripotent embryonic stem cell-like cell.
The new research paper describes how the Stanford scientists focused their research efforts on investigating the role of Oct4, a pluripotency regulator in embryonic stem cells.
To find out what the Oct4 pluripotency regulator does at the one- to two-cell stage, the researchers undertook gene-expression analysis on 42 different genes using Fluidigm's Biomark System for Genetic Analysis, before using Fluidigm's 48.48 dynamic array integrated fluidic circuit to validate differential gene expression between knockdown and control samples.
The Stanford University researchers found that Oct4, the master regulator of embryonic stem-cell pluripotency, also has critical functions during reprogramming of the early mammalian embryo.
The researchers acknowledged how the consistent, high-quality single-embryo data allowed them to identify genes that are consistently differentially regulated and to find rare outlier embryos expressing unique transcriptomes.
Prior to using the Biomark high-throughput system, which can test as little material as a single cell against 96 genes, the group used conventional RT-PCR practices.
The results from these techniques were limited as the methods used samples comprised of pooled cells or embryos, generating relative gene expression that represents an average of all cells assayed.
Additionally, using conventional techniques, the Stanford researchers had not been able to discern between genes that are consistently differentially regulated versus those with a tendency towards stochastic changes or identify rare embryos.
The researchers concluded that what they had accomplished on the Biomark system in three weeks would take more than nine months to accomplish with conventional PCR methods.
