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Culture and Maintenance of Human Embryonic Stem Cells
Published on: December 22, 2009
Defining long-term maintenance conditions of human embryonic stem cells with arrayed cellular microenvironment
David A Brafman1, Kevan D Shah, Thomas Fellner
1Department of Bioengineering, University of California-San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0412, USA.
Stem Cells and Development
|March 31, 2009
Summary
Researchers developed a defined culture system for human embryonic stem cells (hESCs) using array technology. This system optimizes extracellular matrix proteins and signaling molecules for hESC proliferation and self-renewal, paving the way for clinical applications.
Area of Science:
- Stem cell biology
- Biotechnology
Background:
- Clinical applications of human embryonic stem cells (hESCs) require optimized, defined growth conditions.
- Current research primarily focuses on defined media, neglecting defined substrates crucial for hESC proliferation and self-renewal.
- Existing technologies are inadequate for identifying all factors regulating hESC pluripotency and proliferation.
Purpose of the Study:
- To identify fully defined and optimized culture conditions for human embryonic stem cell (hESC) proliferation.
- To develop a defined culture system for the long-term self-renewal of hESCs.
Main Methods:
- Application of multifactorial array technology for systematic screening.
- Screening of extracellular matrix proteins (ECMPs) and signaling molecules.
- Characterization of a completely defined culture system for hESC self-renewal.
Main Results:
- Identification of fully defined culture conditions for hESC proliferation.
- Development and characterization of a defined culture system supporting long-term self-renewal of three independent hESC lines.
- Demonstration of a novel array platform for optimizing stem cell culture.
Conclusions:
- A novel array platform enables the identification of defined culture conditions for hESC proliferation and self-renewal.
- The developed defined culture system supports long-term self-renewal of hESCs.
- Future applications include directed differentiation of hESCs and maintenance of other stem cell populations.
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01:14Maintenance of the ES Cell State
The cells of the blastocyst inner cell mass only remain pluripotent for a short time. This state of pluripotency and self-renewal can be maintained in embryonic stem (ES) cell culture by adding specific chemicals or growth factors to ensure the cells can continue dividing and later differentiate into different cell types. In some cases, the cells are grown on a feeder layer of differentiated cells, which provides the growth factors and extracellular matrix components necessary for stem cell...
00:57Embryonic Stem Cells
Embryonic stem (ES) cells were first discovered in mice in 1981 by Martin Evans. In 1998, James Thomson identified a method to isolate embryonic stem cells from humans. Human embryonic stem cells (hESCs) are obtained from 3-5 day old embryos that remain unused after an in vitro fertilization procedure.
ES cells are grown in a culture medium where they can divide indefinitely, creating ES cell lines. Under certain conditions, ES cells can differentiate, either spontaneously into a variety of...
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