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New iPS cell culture method can reduce the risk of infection caused by transplantation
According to a report on January 13, 2014, Kyoto University in Japan issued a communiqué saying that its research team has developed a new method that can simply cultivate induced pluripotent stem cells (iPS cells) while reducing the risk of infection during transplantation. risk.
Japan’s Kyoto University recently issued a communique stating that its research team has developed a new method that can simply cultivate induced pluripotent stem cells (iPS cells) while reducing the risk of infection during transplantation.
So far, when culturing iPS cells, it is necessary to use a culture medium containing experimental mouse feeder cells and bovine serum to supplement nutrients. However, when using tissues and cells cultured from such iPS cells, animal-derived infections may infect humans, and testing its safety is also time-consuming.
Researchers such as Shinya Yamanaka and Makoto Nakagawa of the Institute of iPS Cells at Kyoto University noticed that a protein called "Laminin-511" can bind cells together, so they used a fragment of this protein "Laminin-511E8" ( LN511E8) produced a culture medium and found that iPS cells can firmly "root" on the culture dish. In order to replace animal ingredients, the research team also produced a culture medium added with amino acids and vitamins, which can increase the number of safer iPS cells.
Since the method developed this time does not use animal ingredients, it can reduce the experimental procedures for confirming safety, is simple and efficient, and is expected to enable iPS cells to achieve clinical applications in the field of regenerative medicine as soon as possible.
The research team also found that the human iPS cells produced by the new method can develop into nerve cells that can produce the neurotransmitter dopamine, cells that can make insulin, and blood cells, which are expected to be used to treat Parkinson's disease and diabetes.
The research team also used this new method to successfully cultivate embryonic stem cells (ES cells) that have the same capabilities as iPS cells. The paper of this achievement has been published in the new issue of British "Scientific Reports" magazine.
Nature Sub-Journal: How to better cultivate iPS cells
On June 26, 2014, although the current culture conditions of human induced pluripotent stem cells (hiPSC) help maintain the pluripotency of the cells, they may also introduce pollutants, which greatly limits the clinical application of these cells. For this reason, the research team of RIKEN in Japan found a better hiPSC training program. They found that CCL2, a chemokine involved in immune response, can replace traditional growth factors in stem cell culture and enhance the pluripotency of stem cells.
Stem cells can differentiate into any cell type in the body. The induced pluripotent stem cell technology developed by Japanese scientist Shinya Yamanaka can reprogram mature cells into pluripotent cells, returning them to a state similar to stem cells and regaining powerful differentiation capabilities. This technology has broad application prospects in the field of regenerative medicine and is regarded as a new hope for cell replacement therapy.
However, although the current culture conditions of human induced pluripotent stem cells (hiPSC) help maintain the pluripotency of the cells, they may also introduce pollutants, which greatly limits the clinical application of these cells.
For this reason, the research team of RIKEN in Japan found a better hiPSC training program. They found that CCL2 , a chemokine involved in immune response, can replace traditional growth factors in stem cell culture and enhance the pluripotency of stem cells. This result was published in Scientific Reports magazine under Nature.
The standard method of culturing hiPSC requires bFGF (basic fibroblast growth factor) to maintain its pluripotent state. Under such culture conditions, hiPSC is actually closer to ectodermal stem cells (epiblast stem cells) rather than true embryonic stem cells.
The researchers replaced the bFGF in the medium with CCL2 for research. They found that CCL2 can also activate the JAK/STAT pathway, which is involved in the immune response and the maintenance of pluripotent stem cells. Studies have shown that culturing with CCL2 enhances the expression of pluripotency marker genes.
In order to fully understand the mechanism of CCL2, the researchers cultured hiPSC with CCL2 and bFGF respectively, and compared their transcriptomes. They found that in stem cells cultured in CCL2, the expression levels of genes related to hypoxia response were higher, such as HIF2A (EPAS1). This shows that CCL2 enhances the pluripotency of stem cells by inducing a response similar to hypoxia. This discovery will attract researchers to further explore the relationship between cell stress (such as hypoxia) and increased cell pluripotency.
Yuki Hasegawa, who led the research, pointed out, “We found that the genes with the most significant differences in expression are related to hypoxia responses. In fact, hypoxia does have an important role in tumor development and maintenance of pluripotency. Our research It can help people further improve the quality of iPSCs and promote the application of these cells in regenerative medicine and disease research."
Studies have also shown that, compared with bFGF, CCL2 cultured hiPSCs have a higher differentiation efficiency. People can use CCL2 to achieve trophoblast-free hiPSC culture to prevent viruses or other pollutants from affecting stem cells. In order to make better use of CCL2 and cultivate more stable human iPSCs, the researchers also developed a special petri dish covered with beads connected to CCL2.
Japan's research and development of new iPS cell culture technology is ten times faster than the current culture
2017-02-10 People's Daily Online, People's Daily Online, Tokyo, February 10, According to the Asahi Shimbun, the research team of Kyoto University and Osaka Gunze Co., Ltd. recently successfully developed a new iPS cell and embryonic stem cell (ES Cell culture technology, the culture speed is ten times the current.
Using iPS cells to make tissues and organs requires culturing a large number of cells. However, the current amount of culture in a petri dish is small, and the efficiency of the method of adding a large amount of culture liquid to the container to stir is relatively low.
The specific associate professor Kenichiro Kamui of the research team and others used gelatin to develop a very fine cloth-like fiber material. By culturing iPS cells in this material, high-quality iPS cells can be cultivated in a short time. This method can shorten the cultivation time to one-fifth to one-tenth of the current one, and can also save costs. Kamui Kenichiro said: "In the future, we will consider increasing the area of this material in order to actually put it into use."
Of course, it is very convenient to reprogram IPS cells.
The experiment involves various experiments such as primary cell culture, electroporation, identification, mycoplasma detection, etc.
The experiments involved mainly include the following product list: Click the blue word to view the details directly