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Printable Handouts
Navigable Slide Index
- Introduction
- Outline
- Pluripotent stem cells (PSC)
- Reprogramming: current methods
- Critical steps in PSC work flow
- Assessing hPSC pluripotency
- Molecular methods
- Expression assay for confirmation of pluripotency
- Two approaches for molecular analysis
- PSC confirmed for pluripotency and differentiation
- Generating optimal gene content
- Testing sensitivity of molecular analysis methods
- Optimal list of genes for focused array
- Workflow
- Reference standards and data analysis
- Sensitivity in detecting differentiation
- Spontaneous differentiation of H9 ESC
- Pluripotency and trilineage differentiation potential
- Sample preparation methods
- Monitoring directed differentiation: neural induction
- Monitoring directed differentiation: mesoderm
- Where can TaqMan hPSC Scorecard be used?
- Summary (1)
- Expression analysis for novel marker identification
- iPSC clones generated using CytoTune
- Clustering of samples based on expression profile
- Identification of CD44 as a differentially expressed
- Confirmation of differential CD44 expression
- CD44 expression in emerging iPSC
- Elimination of CD44+ cells to enhance quality
- Summary (2)
- Acknowledgements
- Information on trademarks
Topics Covered
- Pluripotent stem cells and gene expression analysis
- Plutipotent Stem Cells
- iPSC from various genetic backgrounds using different reprogramming methods and culture conditions
- Need for better standardization methods and markers to confirm cell state
- Gene expression analysis to identify new biomarkers and to confirm cell type based on expression pattern
Talk Citation
Lakshmipathy, U. (2014, March 5). Gene expression analysis of pluripotent stem cells [Video file]. In The Biomedical & Life Sciences Collection, Henry Stewart Talks. Retrieved December 9, 2024, from https://doi.org/10.69645/CAYM9800.Export Citation (RIS)
Publication History
Financial Disclosures
- Dr. Uma Lakshmipathy has not informed HSTalks of any commercial/financial relationship that it is appropriate to disclose.
A selection of talks on Cell Biology
Transcript
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0:00
Hi.
My name is Uma Lakshmipathy.
I'm with the stem cell
R&D at Life Technologies.
My research team is involved in the
development or reprogramming tools
and platforms for stem
cell characterization.
Today, I would like to share
our studies on the use of gene
expression data for the
identification of novel markers
and for rapid characterization and
standardization of pluripotent stem
cells.
0:26
I'll start off with a brief
introduction on pluripotent stem
cells followed by current methods
used to characterize these cells.
I would then like to
share two recent studies.
The first study describes
development of a focus gene
expression panel for
confirming function
and pluripotencies of
ESC and iPSC lines.
This method relies on monitoring
the expression of cell renewal
and lineage markers in
undifferentiated pluripotent cells
and in their corresponding
differentiating cells.
The second study is focused
on identification of CD44
as a negative mark
of pluripotent cells.
1:07
Pluripotent stem cells, or PSCs,
are cells that have the potential
to differentiate into nearly every
type of cell found in the body.
This potential makes them
valuable research tools
for many different applications,
including drug discovery, disease
modeling, and regenerative medicine.
Prior to 2006, the primary
source of pluripotent stem cells
was embryonic stem cells,
or ESCs, which are harvested
from the inner cell mass of a
blastocyst early stage embryo.
In 2006, Shinya
Yamanaka and colleagues
published a landmark paper
describing the derivation
of induced pluripotent stem
cells, or iPSCs, by reprogramming.
Reprogramming is a process
wherein an adult somatic cell,
such as a dermal fibroblast, can
be induced to turn into a cell that
is pluripotent and is essentially
indistinguishable from an ESC.