Introduction: With the iPS research boom driving the rapid development of the entire industry, the iPS tools on the market can be said to be changing with each passing day. iPS cells can theoretically differentiate into any type of cells and have great application prospects in disease research, drug screening and cell therapy. Today, let's take a look at some new products coming out this year.
iPS technology can regain pluripotency of adult cells through reprogramming. iPS cells can theoretically differentiate into any type of cells, and have great application prospects in disease research, drug screening and cell therapy. The iPS research boom has driven the rapid development of the entire industry. The iPS tools on the market can be said to be changing with each passing day. Let us see what new products will be available this year.
Self-replicating RNA
iPS requires the expression of four transcription factors in somatic cells, which together reverse the cell clock to produce embryo-like pluripotent stem cells. In conventional protocols, these factors are delivered by retroviral or lentiviral vectors. Later, people developed new delivery systems such as DNA plasmids, mRNA, and Sendai virus.
However, the above delivery schemes have shortcomings. Retroviruses and lentiviruses are integrated into the host genome. Sendai virus is difficult to remove from culture. Insufficient mRNA requires repeated transfections for several consecutive days. Plasmids may cause harmful genome modifications and are not suitable for some clinical applications.
"The current trend in the iPS space is to replace virus-based delivery strategies with more secure non-viral methods," said Nick Asbrock, product manager at EMDMillipore.
In 2013, Steve Dowdy led the research team at the University of California, developing a "self-replicating RNA" that is as safe as mRNA-mediated reprogramming, but does not require repeated transfection. Based on the VEE virus (Venezuelanequineencephalitis), this RNA encodes an iPS reprogramming factor, a selection marker, and four proteins that help RNA self-replicate. The researchers found that this RNA only needs to be transfected once, enough to induce somatic cells into iPS cells. By simply removing the protein B18R from the medium, these self-replicating RNA molecules can be degraded. The protein B18R is mainly responsible for inhibiting interferon signaling.
EMDMillipore has successfully commercialized this technology and named it Simplicon?. "It provides a more controllable reprogramming path for people without affecting the host's genome," Asbrock explained.
Stemgent has also licensed the technology and will soon launch the relevant reagents, said Brad Hamilton, director of research and development at the company. Stemgent demonstrated at the ISSCR conference last June (International Society for Stem Cell Research) that self-replicating RNA combined with the company's microRNABooster kit can reprogram iPS for blood cells. Blood cells have always been a difficult problem in iPS reprogramming.
In fact, the need for blood cell iPS reprogramming is growing, says Simon Hilcove, product manager at STEMCELL. "The shift from fibroblast reprogramming to blood cell reprogramming is a fairly obvious trend," Hilcove said. "Blood is a more readily available cellular resource that can be easily obtained from patients. There are also a large number of samples stored in the blood bank. In addition, it is generally believed that the genetic risk of blood cells is lower than that of fibroblasts from the skin, because blood cells Not exposed to ultraviolet light (sunlight)."
"The work we showed at the ISSCR was the first to reprogram cells isolated from human blood using RNA technology," Hamilton said. Stemgent currently offers blood cell reprogramming services.
The more perfect medium ThermoFisher is expanding its cell culture products, one of which is CTS?Essential8? MohanVemuri, director of stem cell development at ThermoFisher, said that CTSEssential8 is a fully defined pluripotent stem cell culture medium that is particularly suitable for cell therapy research and applications.
CTSEssential8 medium was developed on the basis of Essential8, which is a pluripotent pluripotent stem cell culture system. “Essential8 has some ingredients derived from human serum,†explains Vemuri. "In CTSEssential8 medium, these components were completely replaced by recombinant proteins."
EMDMillipore's PluriSTEM-XF? Human ES/iPS medium is also one such product. According to Asbrock, this medium can be combined with PluriSTEM-XF recombinant vitronectin (extracellular matrix protein vitronectin) in the absence of MEF (murine embryonic fibroblasts) or tumor extracts (eg BD Matrigel). Support pluripotent cell growth.
These media are all xenobiotic-free formulations that are particularly helpful for preclinical and translational medicine applications. This is because products with uncertain animal sources or chemical compositions can cause regulatory problems and even the risk of introducing unknown pathogens.
More kind of differentiation tool <br> <br> this year, many companies have introduced new products to guide the differentiation of iPS, R & DSystems company StemXVivo? Cardiomyocyte differentiation kit is one of them. "This integrated product can quickly, easily and reproducibly induce human pluripotent stem cells to differentiate into cardiomyocytes," said Julia Hatler, product manager of the company. This kit, launched in September this year, provides a culture medium and a cardiomyocyte-specific antibody that detects differentiation. "With these reagents you can see contracted cells within 11 days," Hatler said.
In addition, R&D Systems has a StemXVivo® kit to induce differentiation of iPS cells into endoderm, ectoderm and mesoderm cells. "These products can promote the differentiation of iPS cells into cells of the three germ layers, and the kit also provides specific antibodies for identifying cell types, and the entire experimental process takes only five days," Hatler said.
R&D Systems' Human Multipotent Stem Cell Function Identification Kit is an intuitive, fast, and low-cost cell pluripotency assessment product. Traditional teratoma experiments are very cumbersome, not only time consuming, but also costly. “Our products are not meant to replace teratoma experiments, but to provide a quick screening solution,†she said.
STEMCELL's new ReproTeSR can reprogram blood cells. Similar to the company's TeSR?-E7?, ReproTeSR "is also non-heterogeneous and produces very well-formed iPS cells that are easy to identify and select," Hilcove said.
Thermo Fisher has also released a new cell differentiation medium. For example, the company's neural induction medium can expand 1 million iPS cells into 20 to 40 million neural stem cells within seven days, Vemuri introduced. Moreover, these cells are highly malleable and are capable of producing precursor cells of the midbrain, hindbrain and forebrain.
Recently, Thermo Fisher has also introduced a medium for differentiating iPS cells into cardiomyocytes. The product can induce human embryonic stem cells or iPS cells to differentiate into a large number of simultaneously beating cardiomyocytes in about ten days.
Despite the emergence of so many new iPS tools this year, the growing demand is still not fully met. For example, researchers also need reliable tools to induce iPS cells into functional islet cells, and Thermo and other companies are working hard to do so. I believe that the iPS tools on the market next year will be more abundant.
iPS technology can regain pluripotency of adult cells through reprogramming. iPS cells can theoretically differentiate into any type of cells, and have great application prospects in disease research, drug screening and cell therapy. The iPS research boom has driven the rapid development of the entire industry. The iPS tools on the market can be said to be changing with each passing day. Let us see what new products will be available this year.
Self-replicating RNA
iPS requires the expression of four transcription factors in somatic cells, which together reverse the cell clock to produce embryo-like pluripotent stem cells. In conventional protocols, these factors are delivered by retroviral or lentiviral vectors. Later, people developed new delivery systems such as DNA plasmids, mRNA, and Sendai virus.
However, the above delivery schemes have shortcomings. Retroviruses and lentiviruses are integrated into the host genome. Sendai virus is difficult to remove from culture. Insufficient mRNA requires repeated transfections for several consecutive days. Plasmids may cause harmful genome modifications and are not suitable for some clinical applications.
"The current trend in the iPS space is to replace virus-based delivery strategies with more secure non-viral methods," said Nick Asbrock, product manager at EMDMillipore.
In 2013, Steve Dowdy led the research team at the University of California, developing a "self-replicating RNA" that is as safe as mRNA-mediated reprogramming, but does not require repeated transfection. Based on the VEE virus (Venezuelanequineencephalitis), this RNA encodes an iPS reprogramming factor, a selection marker, and four proteins that help RNA self-replicate. The researchers found that this RNA only needs to be transfected once, enough to induce somatic cells into iPS cells. By simply removing the protein B18R from the medium, these self-replicating RNA molecules can be degraded. The protein B18R is mainly responsible for inhibiting interferon signaling.
EMDMillipore has successfully commercialized this technology and named it Simplicon?. "It provides a more controllable reprogramming path for people without affecting the host's genome," Asbrock explained.
Stemgent has also licensed the technology and will soon launch the relevant reagents, said Brad Hamilton, director of research and development at the company. Stemgent demonstrated at the ISSCR conference last June (International Society for Stem Cell Research) that self-replicating RNA combined with the company's microRNABooster kit can reprogram iPS for blood cells. Blood cells have always been a difficult problem in iPS reprogramming.
In fact, the need for blood cell iPS reprogramming is growing, says Simon Hilcove, product manager at STEMCELL. "The shift from fibroblast reprogramming to blood cell reprogramming is a fairly obvious trend," Hilcove said. "Blood is a more readily available cellular resource that can be easily obtained from patients. There are also a large number of samples stored in the blood bank. In addition, it is generally believed that the genetic risk of blood cells is lower than that of fibroblasts from the skin, because blood cells Not exposed to ultraviolet light (sunlight)."
"The work we showed at the ISSCR was the first to reprogram cells isolated from human blood using RNA technology," Hamilton said. Stemgent currently offers blood cell reprogramming services.
The more perfect medium ThermoFisher is expanding its cell culture products, one of which is CTS?Essential8? MohanVemuri, director of stem cell development at ThermoFisher, said that CTSEssential8 is a fully defined pluripotent stem cell culture medium that is particularly suitable for cell therapy research and applications.
CTSEssential8 medium was developed on the basis of Essential8, which is a pluripotent pluripotent stem cell culture system. “Essential8 has some ingredients derived from human serum,†explains Vemuri. "In CTSEssential8 medium, these components were completely replaced by recombinant proteins."
EMDMillipore's PluriSTEM-XF? Human ES/iPS medium is also one such product. According to Asbrock, this medium can be combined with PluriSTEM-XF recombinant vitronectin (extracellular matrix protein vitronectin) in the absence of MEF (murine embryonic fibroblasts) or tumor extracts (eg BD Matrigel). Support pluripotent cell growth.
These media are all xenobiotic-free formulations that are particularly helpful for preclinical and translational medicine applications. This is because products with uncertain animal sources or chemical compositions can cause regulatory problems and even the risk of introducing unknown pathogens.
More kind of differentiation tool <br> <br> this year, many companies have introduced new products to guide the differentiation of iPS, R & DSystems company StemXVivo? Cardiomyocyte differentiation kit is one of them. "This integrated product can quickly, easily and reproducibly induce human pluripotent stem cells to differentiate into cardiomyocytes," said Julia Hatler, product manager of the company. This kit, launched in September this year, provides a culture medium and a cardiomyocyte-specific antibody that detects differentiation. "With these reagents you can see contracted cells within 11 days," Hatler said.
In addition, R&D Systems has a StemXVivo® kit to induce differentiation of iPS cells into endoderm, ectoderm and mesoderm cells. "These products can promote the differentiation of iPS cells into cells of the three germ layers, and the kit also provides specific antibodies for identifying cell types, and the entire experimental process takes only five days," Hatler said.
R&D Systems' Human Multipotent Stem Cell Function Identification Kit is an intuitive, fast, and low-cost cell pluripotency assessment product. Traditional teratoma experiments are very cumbersome, not only time consuming, but also costly. “Our products are not meant to replace teratoma experiments, but to provide a quick screening solution,†she said.
STEMCELL's new ReproTeSR can reprogram blood cells. Similar to the company's TeSR?-E7?, ReproTeSR "is also non-heterogeneous and produces very well-formed iPS cells that are easy to identify and select," Hilcove said.
Thermo Fisher has also released a new cell differentiation medium. For example, the company's neural induction medium can expand 1 million iPS cells into 20 to 40 million neural stem cells within seven days, Vemuri introduced. Moreover, these cells are highly malleable and are capable of producing precursor cells of the midbrain, hindbrain and forebrain.
Recently, Thermo Fisher has also introduced a medium for differentiating iPS cells into cardiomyocytes. The product can induce human embryonic stem cells or iPS cells to differentiate into a large number of simultaneously beating cardiomyocytes in about ten days.
Despite the emergence of so many new iPS tools this year, the growing demand is still not fully met. For example, researchers also need reliable tools to induce iPS cells into functional islet cells, and Thermo and other companies are working hard to do so. I believe that the iPS tools on the market next year will be more abundant.
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