Salk scientists and colleagues have proposed new molecular criteria for judging just how close any line of laboratory-generated stem cells comes to mimicking embryonic cells seen in the very earliest stages of human development, known as naïve stem cells.
The tests found that no current protocols lead to truly naïve stem cells, but the guidelines may help researchers achieve that goal by pointing out where each current method falls short. Generating naïve stem cells would be a boon to both basic research and to medical applications of stem cells, such as growing tissue for organ replacement.
"The naïve state potentially has a greater ability to generate different tissue types and could have many uses for regenerative medicine," says senior author Joseph Ecker, professor and director of Salk's Genomic Analysis Laboratory and a Howard Hughes Medical Institute investigator. The work was published online July 14, 2016 in Cell Stem Cell.
While stem cells--cells that have the potential to differentiate into other types of cells--exist in adult humans, the most useful stem cells are those found in embryos, which are pluripotent, capable of becoming nearly any cell in the body. Researchers have developed cocktails of molecules that turn back the clock on adult cells to make them act as stem cells (called induced pluripotent stem cells or iPSCs), and also have cultured lines of stem cells derived directly from embryos (ESCs).
New methods are being formulated to coax the "primed" ESCs--which more resemble cells from post-implantation embryos--back in time even more to resemble naïve stem cells, those found in pre-implantation embryos only days after fertilization. Naïve stem cells are blank slates that form the basis for not only all the cells of the human body, but cells that make up the placenta to support an embryo as well.
"In our opinion, most of the published protocols to generate so-called naïve stem cells are not convincing because they produce cells that are very much like the starting cells--there's not much difference in gene expression," says co-senior author Rudolf Jaenisch of the Whitehead Institute for Biomedical Research and Massachusetts Institute of Technology.
Ecker, working closely with Jaenisch and other collaborators at Whitehead and the Ecole Polytechnique Federale de Lausanne, wanted to see whether these new techniques aiming to induce the naïve stem cell state truly did that. They performed a series of molecular tests on the "primed" cells and ESCs that had been exposed to factors thought to induce naïvety. They estimated the states of the two ESCs by comparing their molecular property and the cells from various stages in embryonic development from previous studies.
Three main tests, they found, were most indicative of the difference between naïve stem cells and other stem cells, letting them place each line of ESCs correctly along this timeline. First, they measured the expression levels of transposons, DNA sequences that can jump around the genome. The expression of certain transposons, they discovered, was indicative of naïve stem cells.
Next, they found that the genomes of naïve embryonic stem cells have less methylation--the addition of methyl chemical groups along DNA. They then studied the state of X chromosomes in naïve cells of female embryos', which each contain two active X chromosomes, unlike more mature embryonic cells that have silenced one X.
Together, the three tests include tens of thousands of genetic biomarkers to characterize the developmental state of stem cells, says co-senior author Didier Trono of the Ecole Polytechnique Federale de Lausanne. "This type of analysis is likely to become a gold standard for quality controlling stem cells, including induced pluripotent stem cells, whether they are to be used solely in research or envisioned for clinical applications," he says.
When current methods for generating naïve stem cells in the lab were judged using the three tests, each fell short of mimicking the naïve embryonic cells in different ways. One new technique, for instance, led to cells that had two active X chromosomes but didn't match the exact methylation patterns desired.
"This really was a comparison of existing methods, applying the same criteria to each method and seeing where each cell state is," says Ecker. "Some of these cells ended up being in states earlier in development and some later in development."
Thorold Theunissen, a postdoctoral fellow in Jaenisch's lab and co-first author of the study, says "Our work provides a rigorous set of criteria for comparing naïve human stem cells to their counterparts in the early human embryo. Previous studies mainly relied on comparisons with mouse stem cells, which are highly divergent from human."
The scientists hope that other research teams adopt their criteria to judge their own methods and cell lines. "The transposon and methylation profiles are pretty standard in terms of technique and protocol so it's pretty easy for other labs to repeat the experiment on, for example, naïve cells from a newly developed method," says Yupeng He, a graduate student in the Ecker lab who helped lead the work.
Other researchers on the study were Styliani Markoulaki, Haoyi Wang, Malkiel A. Cohen, Katherine J. Wert, Yanmei Huang, Jesse Drotar, and Tenzin Lungjangwa of the Whitehead Institute for Biomedical Research; Marc Friedli, Evarist Planet, Julien Pontis, Alexandra Iouranova, Michael Imbeault, and Julien Duc of the Ecole Polytechnique Federale de Lausanne; and Ryan C. O'Neil, Rosa Castanon, Zhuzhu Zhang, and Joseph R. Nery of the Salk Institute.
The work and the researchers were supported by grants from the Simons Foundation, National Institutes of Health, Swiss National Science Foundation, European Research Council, Howard Hughes Medical Institute, Gordon and Betty Moore Foundation, Mary K. Chapman Foundation, a Sir Henry Wellcome Postdoctoral Fellowship, a Foundation Bettencourt Award, the Association Pour La Recherche Sur Le Cancer, and the Fonds de la Recherche en Sante du Quebec.
About the Salk Institute for Biological Studies:
Every cure has a starting point. The Salk Institute embodies Jonas Salk's mission to dare to make dreams into reality. Its internationally renowned and award-winning scientists explore the very foundations of life, seeking new understandings in neuroscience, genetics, immunology and more. The Institute is an independent nonprofit organization and architectural landmark: small by choice, intimate by nature and fearless in the face of any challenge. Be it cancer or Alzheimer's, aging or diabetes, Salk is where cures begin. Learn more at: salk.edu.
Salk Communications | EurekAlert!
Cnidarians remotely control bacteria
21.09.2017 | Christian-Albrechts-Universität zu Kiel
Immune cells may heal bleeding brain after strokes
21.09.2017 | NIH/National Institute of Neurological Disorders and Stroke
Our brains house extremely complex neuronal circuits, whose detailed structures are still largely unknown. This is especially true for the so-called cerebral cortex of mammals, where among other things vision, thoughts or spatial orientation are being computed. Here the rules by which nerve cells are connected to each other are only partly understood. A team of scientists around Moritz Helmstaedter at the Frankfiurt Max Planck Institute for Brain Research and Helene Schmidt (Humboldt University in Berlin) have now discovered a surprisingly precise nerve cell connectivity pattern in the part of the cerebral cortex that is responsible for orienting the individual animal or human in space.
The researchers report online in Nature (Schmidt et al., 2017. Axonal synapse sorting in medial entorhinal cortex, DOI: 10.1038/nature24005) that synapses in...
Whispering gallery mode (WGM) resonators are used to make tiny micro-lasers, sensors, switches, routers and other devices. These tiny structures rely on a...
Using ultrafast flashes of laser and x-ray radiation, scientists at the Max Planck Institute of Quantum Optics (Garching, Germany) took snapshots of the briefest electron motion inside a solid material to date. The electron motion lasted only 750 billionths of the billionth of a second before it fainted, setting a new record of human capability to capture ultrafast processes inside solids!
When x-rays shine onto solid materials or large molecules, an electron is pushed away from its original place near the nucleus of the atom, leaving a hole...
For the first time, physicists have successfully imaged spiral magnetic ordering in a multiferroic material. These materials are considered highly promising candidates for future data storage media. The researchers were able to prove their findings using unique quantum sensors that were developed at Basel University and that can analyze electromagnetic fields on the nanometer scale. The results – obtained by scientists from the University of Basel’s Department of Physics, the Swiss Nanoscience Institute, the University of Montpellier and several laboratories from University Paris-Saclay – were recently published in the journal Nature.
Multiferroics are materials that simultaneously react to electric and magnetic fields. These two properties are rarely found together, and their combined...
MBM ScienceBridge GmbH successfully negotiated a license agreement between University Medical Center Göttingen (UMG) and the biotech company Tissue Systems Holding GmbH about commercial use of a multi-well tissue plate for automated and reliable tissue engineering & drug testing.
MBM ScienceBridge GmbH successfully negotiated a license agreement between University Medical Center Göttingen (UMG) and the biotech company Tissue Systems...
19.09.2017 | Event News
12.09.2017 | Event News
06.09.2017 | Event News
21.09.2017 | Physics and Astronomy
21.09.2017 | Life Sciences
21.09.2017 | Health and Medicine