Acceleration in iPS cells clinical applications!!

[lacazette]

Few other recent advances

-Researchers identify protein crucial for stem cell survival

http://phys.org/news/2015-08-protein-crucial-stem-cell-survival.html#jCp

In a multidisciplinary effort, a team of University of Wisconsin-Madison engineers has identified a protein that is integral to the survival and self-renewal processes of human pluripotent stem cells (hPSC).

The goal for many researchers who work with stem cells is simply to produce an environment that allows the cells to support their own survival. By identifying α-5 laminin as an important key to a cell's survival, the researchers can work to create a synthetic culture environment that encourages the protein's production, says Saha.

Designing new synthetic substrates is about providing a receptive surface that keeps what the cells are producing endogenously in the right place, he says.

"This work gives us a better idea of how to improve the substrate to make a completely defined and inexpensive culture system that's synthetic," says Palecek.


-Researchers develop a method for controlling gene activation (in iPS cells)

http://phys.org/news/2015-09-method-gene.html#jCp

Researchers at the University of Helsinki, Finland, have developed a new method which enables the activation of genes in a cell without changing the genome. Applications of the method include directing the differentiation of stem cells.

The hottest topics in stem cell research at the moment are methods that can regulate the differentiation of cells. The differentiation process is based on how genes in a cell are activated and deactivated, so researchers are looking for ways to control the activation of the genes.

Researchers in Otonkoski's laboratory have now developed a method that enables the regulation of a single gene's behaviour without changing the genome itself. The method employs CRISPR technology, but the regulation itself is controlled by the addition of chemicals. The desired gene is made receptive to the drug by introducing bits of RNA into the cell that will bind to the activator protein and the gene's regulatory area. The gene will then activate in the desired way when the chemicals that regulates the activator protein are provided to the cell.


-A fast and comprehensive method for determining the function of genes could greatly improve our understanding of a wide range of diseases and conditions

http://phys.org/news/2014-01-rewiring-stem-cells.html#nRlv

Mutations with important biological effects can then rapidly be traced to individual genes by next generation DNA sequencing.

"This is a powerful and revolutionary new tool for discovering how gene circuits operate," said Dr Leeb. "The cells and the methodology we've developed could be applied to a huge range of biological questions."




-Hormone found to be critical in promoting growth of human embryonic stem cells, paving way for improved regenerative medicine and cell-based therapies

http://www.a-star.edu.sg/Media/News/Press-Releases/ID/4301/Elabela-identified-as-potential-hormone-for-regenerative-medicine.aspx

Singapore – Scientists from A*STAR’s Institute of Medical Biology (IMB) have discovered that the recently-identified hormone ELABELA is critical in promoting the growth of human embryonic stem cells (hESCs), suggesting its potential as a target for applications in tissue engineering and regenerative medicine.

So far, only a few growth factors for hESCs have been discovered. In a ground-breaking study, IMB has now identified ELABELA as necessary for hESCs to self-renew and differentiate, making it a potential target to stimulate hESC growth, and ensuring its stability for use in regenerative medicine

By activating this pathway, ELABELA also protects the hESCs, and therefore presumably early human embryos, against the intrinsic cell death (apoptosis) pathway which is activated by a variety of cellular and environmental stresses. Given the high susceptibility of hESCs to spontaneous apoptosis and differentiation, ELABELA not only enhances their growth, but also performs the critical function of ensuring their survival.


In regenerative medicine, a key problem is ensuring the stability and survival of hESCs for future differentiation and transplantation. IMB’s discovery implies that clinicians and scientists can target and manipulate ELABELA in order to ensure optimal hESC growth during scale-up of cell production for clinical applications, thus making regenerative medicine cheaper and more accessible, and increasing the chances of successful cell replacement.

[nameless]

http://www.moorfields.nhs.uk/news/new-trial-wet-amd

http://blog.cirm.ca.gov/2015/10/02/stem-cell-stories-that-caught-our-eye-better-heart-muscle-first-patient-with-eye-cell-patch-brain-cross-talk-and-gut-bugs/

London Monday 28 September: A pioneering trial of a new treatment derived from stem cells for people with ‘wet’ age-related macular degeneration (AMD) has commenced at Moorfields Eye Hospital following a successful operation on a patient.

Doctors at the Moorfields Eye Hospital in London have used specialized eye cells derived from embryonic stem cells and grown on a synthetic scaffold to try to reverse blindness caused by age-related macular degeneration(AMD). Prior clinical trials have injected similar cells but without the supporting structure of the patch to hold them in place.

Also, prior trials have aimed to halt the progressive loss of vision in the dry form of macular degeneration. This trial is trying to reverse damage already done by the wet form of AMD. Each of the groups use embryonic stem cells and first mature the cells into a type of cell found in the back of the eye’s retina, retinal pigmented epithelium (RPE) cells.

The first surgery was successfully performed on a patient last month and there have been no complications to date. The patient wishes to remain anonymous, but the team hope to determine her outcome in terms of initial visual recovery by early December (2015).

“The reason we are very excited is that we have been able to create these very specific cells and we have been able to transfer them to the patient,” lead researcher Lyndon Da Cruz told a writer for the Huffington Post. “It’s the combination of being able to create the cells that are missing and demonstrate that we can safely transplant them.”

CIRM funds a team at the University of Southern California and the University of California, Santa Barbara that has collaborated with the London team and plans to use a similar patch system on a trial set to begin in the next few weeks.

“We are delighted to be the site for this very exciting new clinical trial in stem cell therapy, which has the potential to give hope and make such a difference to the lives of people with blinding retinal conditions,”.

https://www.ipscell.com/tag/london-project-to-cure-blindness/

We’ve been hearing a lot this week about an important new clinical study for macular degeneration in the UK. This team is using retinal pigmented epithelial cells (RPEs) made from embryonic stem cells (ESC). They are now testing safety.
There’s real reason for excitement in this area more broadly and that context is important to include, but many newspaper articles haven’t.

For instance, Ocata Therapeutics is doing very similar clinical work and is already years into its FDA-approved clinical trials for macular degeneration using RPEs made from ESC. So far that work has proceeded really well without safety concerns and with hints of efficacy. Also the clinical study in Japan for macular degeneration using IPS cells, while at present on hold, will likely start up again in a new iteration later this year or more likely in early 2016."


it's the same protocol of different hair cure strategies on the making: make the right cells, grow them on a scaffold to then transplant them
so let's all cross our fingers that everything goes well in this trial in term of safety ( with the stemcells post transplant, it's ESCs so it's similar of iPSCs but without the reprogramming process so more safe but still with safety concerns unanswered, and really hope for safety and any kind of problems for using the scaffold thing as we will need that)
Cause if big safety problems comes out, it will add maaany months/years on other regenerative therapies timelines

I think that there is a good chance that there will be safety concerns because they are not just looking for obvious safety issues. They are looking for even a hint of a safety issue. They are looking for even a hint of a theoretical safety issue. I think that the odds are that there will probably be some safety issues.

[lacazette]

Haha you're right that they will not joke with any kind of mini safety issue

but for the moment all things good, this patient is doing well for a month, and the japenese woman with iPSCs passed the one year timeline without any problems, so let's hope it'll continue well like that

http://newsonjapan.com/html/newsdesk/article/113892.php

Patient doing well one year after world's 1st iPS clinical trial
Jiji Press -- Oct 03


And this women was treated with 'dangerous' iPSCs , when they transplanted her in 2014, there wasn't the 100% DNA damage free iPSCs and safe reprogramming process. In fact they didn't even know at that time that the replicative stress during the reprogramming process was leading to small mutations

so i wouldn't be that afraid with safety issues, dna mutations that could lead to cancer is THE safety concern, and is being solved these last months, so Im quite confident personnaly

[FooFighter]

Lacazette, thank you very much for your job here. Its really great, but you are too naive man. Science dont work in that way. For every science breakthrough they need another 10-15 years to proove if treatment is working or not, so this papers are nice, but everything sounds nice on papers. Reality is very different.

[lacazette]

Foo I understand what you mean, but we don' talk about the same kind of papers. You're talking aobut when they announce a precisely big treatment for a type condition,etc.. whereas here it's more various details improvements, ameliorations, technologies discoveries that are making the regenerative medecine clinically applicable

for exemple there were issues for safer reprogramming process, sendaivirus vector was created that lead to free dna damage, and can be purchased now all over the world. There's also now their world wide agreement to sell their combo sendaivirus/naive state that offer better benefit. and another company sell safe reprogramming process protocols with the use of RNA that it seems to be even more powerful

So does it took here 10/15 years between discovery and practical use? no, the majority of papers here are achievements for technology/knowledge improvement to make things cheaper, faster, easier, safer on the iPSCs treatments area. When these kind of breaktroughs comes out it's a matter of months or even days for other researchers to use it or replicate the protocol
we don't talk here about a treatment that has to be proven as you mentioned

[FooFighter]

Lacazette, i can tell you that a lot of scientist and companies lie about their papers and products just to get public attention and money. How we will know that some paper from scientist work on human? How we will know that somenthing publiced is not a lie?

[JayM]

Lacazette, i can tell you that a lot of scientist and companies lie about their papers and products just to get public attention and money. How we will know that some paper from scientist work on human? How we will know that somenthing publiced is not a lie?

So does it took here 10/15 years between discovery and practical use? no, the majority of papers here are achievements for technology/knowledge improvement to make things cheaper, faster, easier, safer on the iPSCs treatments area. When these kind of breaktroughs comes out it's a matter of months or even days for other researchers to use it or replicate the protocol
we don't talk here about a treatment that has to be proven as you mentioned

Come on dude are you even reading what he said?

[lacazette]

Last publish from the inventor of iPSCs, confirming the recent progress on reprogramming safety and efficiency

2015 Oct 6
Practical Integration-Free Episomal Methods for Generating Human Induced Pluripotent Stem Cells.

The advent of induced pluripotent stem (iPS) cell technology has revolutionized biomedicine and basic research by yielding cells with embryonic stem (ES) cell-like properties. The use of iPS-derived cells for cell-based therapies and modeling of human disease holds great potential. While the initial description of iPS cells involved overexpression of four transcription factors via viral vectors that integrated within genomic DNA, advances in recent years by our group and others have led to safer and higher quality iPS cells with greater efficiency. Here, we describe commonly practiced methods for non-integrating induced pluripotent stem cell generation using nucleofection of episomal reprogramming plasmids. These methods are adapted from recent studies that demonstrate increased hiPS cell reprogramming efficacy with the application of three powerful episomal hiPS cell reprogramming factor vectors and the inclusion of an accessory vector expressing EBNA1

http://www.ncbi.nlm.nih.gov/pubmed/26439714

[joachim]

Last publish from the inventor of iPSCs, confirming the recent progress on reprogramming safety and efficiency

2015 Oct 6
Practical Integration-Free Episomal Methods for Generating Human Induced Pluripotent Stem Cells.

The advent of induced pluripotent stem (iPS) cell technology has revolutionized biomedicine and basic research by yielding cells with embryonic stem (ES) cell-like properties. The use of iPS-derived cells for cell-based therapies and modeling of human disease holds great potential. While the initial description of iPS cells involved overexpression of four transcription factors via viral vectors that integrated within genomic DNA, advances in recent years by our group and others have led to safer and higher quality iPS cells with greater efficiency. Here, we describe commonly practiced methods for non-integrating induced pluripotent stem cell generation using nucleofection of episomal reprogramming plasmids. These methods are adapted from recent studies that demonstrate increased hiPS cell reprogramming efficacy with the application of three powerful episomal hiPS cell reprogramming factor vectors and the inclusion of an accessory vector expressing EBNA1

http://www.ncbi.nlm.nih.gov/pubmed/26439714

fantastic to see how fast they are progressing with iPS. the discovery of iPS is really a breakthrough for mankind and future modern medicine. together with the new CRISPR-cas9 and cpf1 gene editing methods, those two tools will cure every damn disease longterm.

[lacazette]

still few ipsc things

-21 sept 2015 New Method for Testing Induced Pluripotent Stem Cells Differentiation Potential Could Lead to Safer and More Potent Treatments

http://www.streetinsider.com/Press+Releases/New+Method+for+Testing+Induced+Pluripotent+Stem+Cells+Differentiation+Potential+Could+Lead+to+Safer+and+More+Potent+Treatments/10906066.html

During recent collaboration, two companies, Atlas Regeneration and Insilico Medicine, demonstrated the close resemblance of iPSCs with ESCs at the pathway level, and provided examples of how pathway activity analysis can be applied to identify iPSC line abnormalities or to predict in vitro differentiation potential. The results indicate that pathway activation profiling is a promising strategy for evaluating the safety and potency of iPSC lines in translational medicine applications allowing scientists to test differentiation abilities of many iPSC lines in silico while saving valuable time for patients waiting for treatment.

"Regeneration Intelligence is unique among pathway analysis platforms. Using our algorithm along with proprietary pathway database, we established for the first time pathway activation profiles of iPS.

Anthony Atala, MD, CEO of Atlas Regeneration said, "Our Regeneration Intelligence platform has been used in many iPSC lines and is helping stem cells biologist to improve and speed up decision-making. Unfortunately, the entire process of verification and validation of differentiation abilities using in vitro differentiation assays typically takes 12 weeks and time is critical for definitive treatment, especially in urgent cases. With the help of Regeneration Intelligence, we may be able to significantly reduce the time and cost of the process."


-Bromodeoxyuridine promotes full-chemical induction of mouse pluripotent stem cells

http://www.nature.com/cr/journal/v25/n10/full/cr201596a.html

Direct reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) with transcription factors (e.g., Oct4 (O), Sox2 (S), Klf4 (K), and c-Myc (M)) greatly expands our understanding of cell fate control. iPSCs resemble embryonic stem cells (ESCs) but without immune rejection and ethic issues, and are therefore considered as a promising source for cell replacement therapy.

However, iPSC applications are hindered by safety concerns about the possible genetic alterations caused by the use of exogenous pluripotency-associated factors. Many efforts have been taken to make iPSCs more amendable in clinical applications by using non-integrating gene delivery approaches2, or cell membrane-permeable proteins3,4 to induce the reprogramming.

Small-molecule compounds have also been found to be extremely useful in facilitating iPSC generation and can replace several reprogramming factors5. Several combinations of small-molecule compounds have been reported to allow iPSC generation with only Oct46,7. However, complete chemical-mediated reprogramming of somatic cells into the pluripotent state has been proved to be extremely difficult.

Here we report that the commonly used biological reagent, bromodeoxyuridine (BrdU), is able to enhance Yamanaka factor-mediated reprogramming. More interestingly, BrdU can replace Oct4, the most critical factor in iPSC generation. Further studies demonstrate that BrdU promotes full-chemical induction of mouse iPSCs using several chemical ****tails, with the minimal combination being BrdU, CHIR99021, Repsox, and Forskolin. These iPSCs resemble ESCs in terms of their gene expression, epigenetic status, in vivo differentiation potentials and the ability to generate chimera

In summary, we demonstrate that BrdU can replace Oct4, the most critical factor in iPSC generation, and promotes full-chemical induction of mouse iPSCs with the minimal combination being BrdU, CHIR99021, Repsox and Forskolin. Since BrdU has already been used in patients12, this combination may lay a foundation for full-chemical induction of human iPSCs and may eventually provide a safer strategy to generate clinically applicable iPSCs.


-Enhanced mRNA reprogramming by Reprocell

https://www.reprocell.com/en/products/research/reagent-reprog/micro-rna-reprog

The Stemgent mRNA Reprogramming Kit is the fastest, safest, and most efficient method for generating integration-free, virus-free, clinically relevant human iPS cells.

The total time needed to generate a characterized iPS cell line using virus-based systems can take up to 25 weeks, whereas the Stemgent mRNA Reprogramming System generates virus-free, integration-free iPS cell lines in less than 2 weeks. This system enables the generation of fully characterized and banked iPS cell lines ready to use in as little as 9 weeks

The Stemgent mRNA Reprogramming System provides efficiencies greater than 1% as compared to other methods, which yield reprogramming efficiencies varying from 0.00001 to 0.01%. In addition to increased yield of colonies and fast reprogramming kinetics, mRNA reprogramming does not require laborious multi-step passaging or screening for viral or genomic integration once the new colonies are derived.
The mRNA Reprogramming System eliminates virus bio-containment and safety issues, and carries no risk for insertional mutagenesis, an inherent concern with DNA-based reprogramming methods.


-the Top 5 iPS Cell Influencers : https://www.bioinformant.com/do-you-...ers-right-now/

[ShookOnes]

I think what Foofighters is trying to say is that with all these new papers you've been sharing, the real possible cure is 10-15 years away with these current findings. ( I think...and 10-15 imo is fairly highly optimistic)..

[lacazette]

A quote before the summer from S&B and european institute researchers:


"In order to obtain cells suitable for clinical application, transgene-free iPSCs need to be generated to avoid transgene reactivation, altered gene expression and misguided differentiation. Moreover, a highly efficient and inexpensive reprogramming method is necessary to derive sufficient iPSCs for therapeutic purposes. "

http://www.jove.com/video/52885/generation-induced-pluripotent-stem-cells-from-frozen-buffy-coats

-Protocols for transgene-free iPSCs are now reality and being sell by leading companies in this domain


-And the second problem will soon be something of the past,

with things like the Automation of iPS cell lines production by the NYSCF robot we already talked here (http://stemcellassays.com/2015/10/ips-automation/)

or with this :

18sept2015

http://www.stem-cells-news.com/1/researchers-improve-method-to-create-induced-pluripotent-stem-cells/

University of Minnesota Medical School researchers have developed a new strategy to improve the development of induced pluripotent stem cells (iPS).

Currently, iPS cells are created by introducing four defined genes to an adult cell. The genes reprogram the adult cell into a stem cell, which can differentiate into many different types of the cells in the body. Typically, the four genes introduced are Oct4, Sox2, Klf4 and c-Myc, a combination known as OSKM.

The U of M researchers found that by fusing two proteins – a master stem cell regulator (Oct4) and a fragment of a muscle cell inducer (MyoD) – they succeeded in “powering up” the stem cell regulator, which can dramatically improve the efficiency and purity of reprogrammed iPS cells.

“Our team discovered that by fusing a fragment of the powerful protein MyoD to Oct4 we could create a ‘super gene’ which would improve the iPS reprogramming process,” said senior author Dr. Nobuaki Kikyo, Stem Cell Institute researcher and University of Minnesota Medical School associate professor. “The result is what we termed M3O, or ‘super Oct4’ – a gene that improves the creation of iPS cells in a number of ways. In the process we shed new light on the mechanism of making iPS cells.”

The challenge with the previous method – OSKM – has been that very few cells actually become iPS cells during reprogramming. In fact, the rates currently stand at about 0.1 percent. Another issue has been tumor development. Because some of the reprogramming genes introduced are oncogenes, the risk of developing tumors grows.

The research, led by Kikyo and Dr. Hiroyuki Hirai, both from University of Minnesota Medical School and Stem Cell Institute, led to a new gene model that minimizes such complications while amplifying the benefits of the process.

According to Kikyo, the new gene model – called M3O-SKM – improves iPS development by:

Increasing efficiency. The efficiency of making mouse and human iPS cells was increased over 50-fold compared with the standard OSKM combination.
Increasing purity. The purity of the iPS cells was much higher with the M3O-SKM gene introduction (98% of the colonies) compared with OSKM (5%).
Facilitating the reprogramming. iPS cell colonies appeared in around five days with M3O-SKM, in contrast to around two weeks with OSKM.
Decreasing the potential for tumor formation. M3O achieved high efficiency of making iPS cells without c-Myc, an oncogene that can potentially lead to tumor formation.


In addition, human iPS cells usually require co-culture with feeder cells typically prepared from mouse cells, obviously creating a problem when the cells are destined for human transplantation.[COLOR="red"] The M3O model did not require such feeder cells, greatly simplifying the process.

Future Impact

According to senior author Kikyo, this new strategy will dramatically speed up the process of making patient-specific iPS cells, which makes clinical applications via transplantation of the cells more feasible to treat many diseases incurable otherwise."""




terskikh, tsuji, ohyama, Fujiwara and others need these efficient iPSC reprogramming progresses to have a chance to test their technique on human.
But what is good is that these tools are needed by any researchers/companies in the world for ANY diseases potentially treated with iPSCs, that's why it's going fast til this summer
According to researchers it is the hot topic of the moment in regenerative medecine as a safe and efficient iPSCs reprog protocol will open doors to clinical applications


And as you see above they are finding great things.

In the same articile they talk also about a new area: direct reprogramming !!


Many researchers are also examining how to reprogram one cell type into another without going through iPS cells; for instance, coaxing skin cells into becoming neurons or pancreas cells by introducing several genes.

The approach, called direct reprogramming, is thought to be the next generation approach beyond iPS cell technology.

The U of M approach – fusing a powerful protein fragment to other host proteins – can be widely applied to the direct reprogramming approach as well



We could maybe soon ear about adult skin cells that are directly reprogrammed in DPs cells without the iPSCs transition

[JayM]

Lacazette have you read this - http://news.xinhuanet.com/english/20..._134748306.htm

Could be big news for hair because making blood vessels is needed if you want large scale skin grafts ect and better models.

[lacazette]

Hey jay yeah that's really huge! it's crazy the solutions we will have in 20/30 years when a lot of us will be dealing with serious diseases, it's gonna be insane! BUT we need hair ASAP to enjoy this fukcing life now! hehe

[lacazette]

Big progress are coming out day after day, and few days ago the 2 most powerful domains in regenerative medecine are now meeting eachother : they are now able to bioprint iPS cells!
( safety, time consuming, cheapness, easiness, make large amounts of, etc,etc the meeting of these two worlds will be a huge help in every areas needed for commercialisation be possible one day)


21oct

http://3dprintingindustry.com/2015/10/21/scottish-university-engineer-creates-bioprinter-capable-of-3d-printing-stem-cells/

-------
In a new breakthrough for bioengineered 3D printing, a team from the Scottish-based Heriot-Watt University’s School of Physical Sciences and Engineering has constructed a 3D printer that is able to print with delicate stem cell cultures
The printer is engineered to print what the team calls ‘induced pluripotent stem’ (iPS) cells, which are delicate cells derived from the particular donor.

“This study is the first to demonstrate that human induced pluripotent stem cells, that is stem cells derived from the adult patient’s own cells, can be bioprinted without adversely affecting their biological functions; that our 3D printing process is gentle enough to do this,” said Dr. Shu

Dr. Shu is describing a system that is able to bioprint these sensitive cell cultures without destroying their biological function to create different cell types.

“The ability to bioprint stem cells while either maintaining their pluripotency, their ability to develop into all types of cells in the body, or indeed directing their differentiation into specific cell types, will pave the way for producing organoids, or tissues on demand, from patient specific cells,” Dr. Shu added.
------


imagine,guys like Terskikh had to take adult cells from a biopsie, make them pluripotent, and then make these iPSCs become DPs cells , all that manually . It's commercially not possible for a treatment when you need many many DPs cells. But now the entire process could be done via the bioprinter
Not only transform the adult cells into iPSCs, but also then directing those ones into the type of cells needed

Complete safety reprogramming is currently being solved, now that mechanical automation via bioprinting will enter the game , viability of clinical application of a treatment is becoming more and more possible and concrete day after day

I still maintain that we will see at least one of them enter a phase1 during 2016 (or at least a proof of concept on a human head) ( terskikh, takashi tsuji, regience, ohyama, amagai, CHA korea, P&G/Singapore, chineses ones, or another using iPSC technology ) ( I could add also Cristiano as I see she is presenting recently a " functional complex skin derived from iPSC" )