Out of 8 embryo cells: if 1 turned to an ES cell, 7 could still become a child

At least I know how to start my stem cell comprehensive exam tomorrow (The trick is to use blastocyst medium supplemented with laminin and fibronectin):

Human Embryonic Stem Cell Lines Generated without Embryo Destruction

Young Chung, Irina Klimanskaya, Sandy Becker, Tong Li, Marc Maserati, Shi-Jiang Lu, Tamara Zdravkovic, Dusko Ilic, Olga Genbacev, Susan Fisher, Ana Krtolica, and Robert Lanza

“To date, the derivation of all human embryonic stem cell (hESC) lines has involved destruction of embryos. We previously demonstrated that hESCs can be generated from single blastomeres (Klimanskaya et al., 2006). In that ‘‘proof-of-principle’’ study, multiple cells were removed from each embryo and none of the embryos were allowed to continue development. Here we report the derivation of five hESC lines without embryo destruction, including one without hESC coculture. Single blastomeres were removed from the embryos by using a technique similar to preimplantation genetic diagnosis (PGD). The biopsied embryos were grown to the blastocyst stage and frozen. The blastomeres were cultured by using a modified approach aimed at recreating the ICM niche, which substantially improved the efficiency of the hESC derivation to rates comparable to whole embryo derivations. All five lines maintained normal karyotype and markers of pluripotency for up to more than 50 passages and differentiated into all three germ layers.”

Legend: “(A) Stages of derivation of hES cells from single blastomere. (a) Blastomere biopsy, (b) biopsied blastomere (arrow) and parent embryo are developing next to each other, (c) initial outgrowth of single blastomere on MEFs, 6 days, and (d) colony of single blastomere-derived hES cells.”

Context link: Wired: Embryonic Stem Cells Created Without Harming Embryo, for Real This Time

California embryonic stem cell dollars: beyond the embryonic phase

According to the newest Request For Applications (RFA) of the California Institute for Regenerative Medicine (CIRM), the New Cell Line Awards will support two categories of research:

Category 1: Derivation of new hESC lines using excess or rejected early stage human
embryos generated by in vitro fertilization.

Category 2: Derivation of pluripotent human stem cell lines from other sources using
alternative methods such as (but not limited to) SCNT or reprogramming of neonatal or
adult cells (iPS cells).

The real news is encoded in category 2: from now on even adult stem cell research can be backed by California Embryonic Stem Cell Dollars. The same idea in another form in the text:

• disease-specific or otherwise genetically diverse, pluripotent stem cell lines to support
studying the effects of genetic variation on disease mechanism and response to
treatment, and the discovery and evaluation of new drug candidates
• the discovery and implementation of alternative methods for generating pluripotent
human stem cells, including technology leading to the generation of patient-matched or
disease-specific cell lines

What research trend is behind? The generation of induced pluripotent stem (iPS) cells. The successful reprogramming of differentiated human somatic cells into a pluripotent, embryonic stem (ES) cell-like state that would allow creation of patient- and disease-specific stem cells instead of using controversial embryonic stem cells was recently reported by 2 groups of researchers: the Yamanaka and the Thomson lab.

Under this RFA, CIRM intends to commit up to $25 million to support up to 16 awards,
eight ( in each of the two categories of research. CIRM proposes to fund each award
for up to three (3) years for direct project costs of up to $300,000 per year.

Other side of the story in California Stem Cell Report: California’s Widening Stem Cell Net

Unofficial and hypercritical peer review of a paper on p53’s role in hESC regulation

Once I wrote shortly about the following peer review paper which was popped out of my PubMed feeds to draw some attention to it: Han Qin, Tianxin Yu, Tingting Qing, Yanxia Liu, Yang Zhao, Jun Cai, Jian Li, Zhihua Song, Xiuxia Qu, Peng Zhou, Jiong Wu, Mingxiao Ding, Hongkui Deng
Regulation of apoptosis and differentiation by p53 in human embryonic stem cells.
J Biol Chem. 2007 Feb 23;282(8):5842-52 doi:10.1074/jbc.M610464200

Now I got a very thorough comment on this paper by an author nicknamed “wolf” which systematically goes through the paper and gives a highly critical peer review of it. So I just publish the comments and next ask the authors (the first or the last) of the criticized paper and give them the possibility to defend their experiments and statements. My role here is the role of the blog”publisher”.

Comments re Qin paper p53 and hESC apoptosis by “wolf”:

This paper starts with making a fundamental mistake in not determining the kinetics of UV induced apoptosis and therefore missing the modulation of p53 target genes.
Subsequently they attempt to explain the absence of this by using transient transfections and analysing the cells at timepoints when half of the cells (mainly the undiff cells) are already dead and then interpret the data of the differentiated transfected (more resistant) hESC as if they were undiff hESC. The paper then desperately tries to come up with explanations for their own contradictory results). The data set further lacks controls (lentiviral mock transduced cells, no isotype controls etc), uses the wrong assays (such as PI staining to assess apoptosis, morphological assessment of differentiation by surface area) and lacks insight into the mechanisms controlling apoptosis (no cyt c release, no idea how p53 by itself might trigger mitochondrial apoptosis, etc).

Specifically;

Materials and Methods

Page 2: The authors use mainly one line of late passage hESC (p42-p6 grown in KSOR, which are highly CD30 positive leading to alterations in apoptosis regulation. We use three hESC lines at passages before p12 only.

Page 3: endoderm differentiation occurs in 4 days after Activin addition ? This is very quick with >80 % of hESC expressing sox17 after 100 ng/ml activin ?

Page 3: In immunostaining no antibody controls were used instead of isotype control with identical concentrations. We use isotype controls for all our immunos.

The first stem cell related Nobel prize: Martin Evans, 2007

The Nobel Assembly at Karolinska Institute has today decided to award The Nobel Prize in Physiology or Medicine for 2007 jointly to Mario R. Capecchi, Martin J. Evans and Oliver Smithies for their discoveries of “principles for introducing specific gene modifications in mice by the use of embryonic stem cells” Link

It’s rather a 2/3 gene technology, 1/3 stem cell methodology Prize, but shows how those technologies are interrelated.

Evans is still an active scientist but here are 2 famous papers he authored or coauthored:

1. First teratocarcinoma paper (always those cancer cells), how to isolate and maintain them, Evans is the single author: The isolation and properties of a clonal tissue culture strain of pluripotent mouse teratoma cells. J Embryol Exp Morphol. 1972 Aug;28(1):163-76.

2. And then the switch to embryonic stem cells a decade later, a protocol lead to the 1998 Science paper by Thomson et al. on human ES cells: Evans, M. J. & Kaufman, M. H. Establishment in culture of pluripotential cells from mouse embryos. Nature 292, 154-6. (1981).

Vincent van Gogh and human embryonic stem cells

Let’s continue our Hit art illustrations for scientific slides project this time with van Gogh’s “Starry night“. The slide is from Chang-Kyu Lee’s presentation on the SENS3 conference, entitled Interspecies somatic cell nuclear transfer for establishing embryonic stem cells with desired genotype.

George Daley explains the source of Hwang’s “cloned” ES cells on Youtube

George Daley, the new president of the International Society for Stem Cell Research explains shortly the notorious case on a not embeddable (??????) YouTube video. If you are too busy to read the story, than watch it, it is 2 minutes and 13 seconds. Thanks for the video tip, Alexey Bersenev.

If you have a bit more time to read on:

Hwang’s “clone” was really a parthenote, Daley reports

Donating Frozen Embryos for Stem Cell Research: a survey in Science

EMBRYONIC STEM CELLS: Willingness to Donate Frozen Embryos for Stem Cell Research by Anne Drapkin Lyerly and Ruth R. Faden, Science 6 July 2007: Vol. 317. no. 5834, pp. 46 - 47 DOI: 10.1126/science.1145067

We conducted a survey of 2210 infertility patients receiving treatment at one of nine major, geographically diverse infertility centers and asked these patients about their intentions for the embryos they currently stored. Participating centers were located in California, Colorado, the District of Columbia, Maryland, Missouri, New Jersey, North Carolina, Oregon, Pennsylvania, and Virginia. The respondents were asked to answer a set of questions with one of the following: very likely, somewhat likely, somewhat unlikely, very unlikely, or unsure/don’t know.

/Figure legend. Disposition option for some or all of cryopreserved embryos currently stored. SCNT, somatic cell nuclear transfer. Key: Somewhat likely (lavender), very likely (gray).
CREDIT: MIODRAG STOJKOVIC/SCIENCE PHOTO LIBRARY /

Prepare for the ISSCR 2007 stem cell party! June, 17-20

5th International Society for Stem Cell Research (ISCCR) Annual Meeting will be held this weekend June 17-20, at the Cairns Convention Centre Queensland, Australia and the schedule is exciting. Unfortunately it is improbable that there will be a pioneer web coverage on this mainstream congress as it happened in the case of the Edmonton Aging Symposium which According to the organizers “was a WORLD FIRST! in being streamed live onto the internet and indeed there was a full video, audio and presentation access. Anyhow, here are 2 more interesting shots from the ISSCR schedule:

Nature Reports Stem Cells is live from now on…for stem cell enthusiasts

No more waiting: Nature Reports Stem Cells (NRSC) launched today, and so finally there is a fully web native, scientifically high-end (naturally), freely accessible, all-in-one stem cell research hub site for everyone (especially for the researchers) to read, share, contribute and turn the acquired insights back into new experiments, policies, ethics, businesses and clinical trials.

Edited by the devoted small team Natalie DeWitt (Editor at Large), Monya Baker (News Editor) and Jessica Kolman (Editorial Assistant), based in Nature’s San Francisco office, California (where else?) NRSC has a bunch of usual and unusual ways: news, featured editor, journal club with user recommended articles and voting system, interviews, events and the really exciting and experimental Insider the paper section. From the first editorial of NRSC: “Inside the Paper posts edited discussions between authors and reviewers during peer review. Such transparency should not only deepen readers’ understandings of individual research publications, it will expose the workings of peer review itself. In the coming months, we plan to launch a Toolbox section will aggregate information on stem-cell protocols, reagents, and cell lines that would otherwise require trawling through literature or having serendipitous conversations at conferences.”

On CIRM’s Medical and Ethical Standards in Plos Medicine

Geoffrey P. Lomax, Zach W. Hall, Bernard Lo: Responsible Oversight of Human Stem Cell Research: The California Institute for Regenerative Medicine’s Medical and Ethical Standards

Source: Plos blog, In the May issue of PLoS Medicine

Anthony Atala on regenerative medicine at New Yorker’s 2012 conference

Dr. Anthony Atala of pluripotent amniotic fluid-derived stem cell and tissue engineered bladder fame gave a presentation on Regenerative Medicine at the 2007 New Yorker Conference “2012: Stories from the Near Future”.

Atala, the director of the Biopolis-like huge Wake Forest Institute for Regenerative Medicine with circa 150 people, talks amongst others on the differences amongst embryonic, placental and adult stem cells, the differences between the tissue engineering of solid organs (kidney) and hollow organs (bladder) and on the five-year follow-up careful strategy behind the successful tissue engineered products. Here are some slides of the presentation in the order of appearance:

Stem Cell Cup: California Embryonics vs Massachusetts Oldies: $3 vs $1.25 Billion

While people in California can think they are in the centre of the (embryonic) stem cell universe due to Proposition 71’s $3 Billion and the invasion of good scientists into the West Coast, Massachusetts academic and biotech people also are thinking along those lines, so state officials quickly set up a $1.25 Billion package for funding stem cell research in the very state and “establishing the first stem cell bank, a repository of all the stem cell lines created in Massachusetts laboratories, which would serve as a kind of stem cell lending library to scientists around the world. “

Links:

Massachusetts floats $1 bln stem cell research plan (Reuters)

Massachusetts Proposes Stem Cell Research Grants (New York Times)

2 models of embryonic and adult blood formation: Figure by Ueno and Weissman

If you have previously thought (in your spare time) that the conventional wisdom concerning blood formation is that the yolk sac’s embryonic blood-forming cells serve only the embryo, while the source of adult blood-forming stem cells is the region called aorta-gonad-mesonephros (AGM), it’s time to think it again due the elegant experiments of Samokhalov et al.: Cell tracing shows the contribution of the yolk sac to adult haematopoiesis Nature 446, 1056-1061 (26 April 2007)

Legend: a, The ’separate’ model.

Stem Cell Research Budget Cutting in Florida

It is just $20 million but look at the policy behind: Stem cell research faces budget shortfall