12 results about "Mesoderm Cell" patented technology

The application provides a freezing and recovery method and application of pluripotent stem cell induced differentiation middle stage cells, and the freezing method comprises the following steps: (1) the starting cells are pluripotent stem cells, which are induced to the middle stage; (2) the cells are recovered and counted by using a mild digestion enzyme, and the cells are resuspended in a freezing solution; (3) after programmed cooling to-80 DEG C, the cells are transferred into liquid nitrogen for storage, and are ready for use. The freezing method and the freezing solution provided by the application significantly improve the freezing survival rate of the differentiation middle stage cells, avoid the apoptosis or irreversible stress damage induced by the traditional freezing solution, and obviously improve the problems of cell damage and low survival rate caused by the general freezing solution in the prior art. The differentiation potential of the recovered cells is good, and the cells can successfully form mature kidney unit structures, so that the problem of subsequent differentiation failure in the prior art is solved.

Disclosed herein are improved in vitro methods of making splanchnic mesoderm cell types and subtypes thereof from pluripotent cells. These methods can be used to produced improved foregut- and hindgut-derived organoids containing enriched mesenchyme, which enhances organoid viability, growth, and maturation, both in in vitro culture and in vivo transplantation.

The invention discloses a vascular endothelial cell as well as a preparation method and application thereof. The preparation method comprises the following steps: inoculating induced pluripotent stem cells into a culture medium containing an ROCK signal channel inhibitor for culturing, and inducing the cultured induced pluripotent stem cells by using a culture medium containing a Wnt signal channel activator and a BMP signal channel activator, so that the induced pluripotent stem cells are differentiated into mesoderm cells; inducing the mesoderm cells by using a culture medium containing a vascular endothelial growth factor and a cAMP signaling pathway agonist, so that the mesoderm cells are differentiated into endothelial coarse cells; and performing positive sorting of the endothelial cell marker CD144 on the endothelial coarse cells to obtain purified vascular endothelial cells. The induction path of the method is clear and efficient, the period is short, the vascular endothelial cell population with high purity, complete vascular barrier function and mature phenotype can be obtained, and the cell population can be applied to construction of a vascular network in a gel matrix.

The invention discloses a kit and a method for constructing a human bone organoid from pluripotent stem cells, and relates to the technical field of organoid culture. The kit for constructing the human bone organoid by the pluripotent stem cells comprises a culture medium A, a culture medium B, a culture medium C, a culture medium D and a culture medium E. The culture medium A induces the pluripotent stem cells to form original stripes, the culture medium B induces the original stripes to form paraaxial mesoderm, the culture medium C induces the paraaxial mesoderm to form body wall mesoderm, the culture medium D induces body wall mesoderm cells to differentiate to bone cells, and the culture medium E induces bone cells to form cartilage organs. The kit provided by the invention can induce the pluripotent stem cells into bone organs, and has important significance in research and treatment of pathogenesis of bone related diseases and related research of bone repair and regeneration.

Disclosed herein are improved in vitro methods of making splanchnic mesoderm cell types and subtypes thereof from pluripotent cells. These methods can be used to produced improved foregut- and hindgut-derived organoids containing enriched mesenchyme, which enhances organoid viability, growth, and maturation, both in in vitro culture and in vivo transplantation.

The invention discloses a method for obtaining, activating and amplifying natural killer cells through iPSC differentiation induction, and relates to the technical field of stem cell biology, the method for obtaining, activating and amplifying natural killer cells through iPSC differentiation induction comprises the following steps: S1, preparing and inoculating iPSC cells; s2, differentiating the iPSC to a mesoderm; s3, the mesoderm cells are differentiated to CD34 + hematopoietic endothelial cells; s4, differentiating the CD43 < + > / CD45 < + > hematopoietic stem cells into the iNK cells; s5, maturation of the iNK cells; s6, activating and amplifying the iNK cells; in the step S1, cells are resuspended by using a fresh E8 complete culture medium and are inoculated into a porous culture plate; a plurality of induction culture media are used in the culture process; by adopting a porous culture plate and using an E8 complete culture medium to resuspend cells, efficient inoculation and primary culture of iPSC are realized, and the survival rate and amplification efficiency of cells are improved; the differentiation process of the iPSC towards the mesoderm, the hematopoietic endothelial cells and the CD34 + hematopoietic stem cells is optimized by using various induction culture media, and the differentiation efficiency is improved.

The invention relates to a method for producing a mesenchymal stem cell (MSC), the method comprising culturing a primitive mesoderm cell in a mesenchymal colony forming medium (M-CFM) comprising LiCl and FGF2, but excluding PDGF, under normoxic conditions for sufficient time for a mesenchymal colony to form, and culturing the mesenchymal colony adherently to produce the MSC, wherein the MSC has superior T-cell immunosuppressive properties relative to an MSC not produced in said M-CFM. The invention also relates to an MSC produced by the method, a population of MSCs produced by the method, a therapeutic composition comprising the MSC produced by the method, an M-CFM and an M-CFM in concentrated form, and method and uses of the MSC or population in treating a disease.

The invention relates to a method for producing mesenchymal stem cells with bone differentiation tendency by using extraembryonic mesoderm cells. Specifically, the invention develops a method for inducing and differentiating human pluripotent stem cells into embryonic ectomesoderm cells in vitro, and optimizes induction culture conditions and a purification method. The obtained extraembryonic mesoderm cells are further subjected to differentiation culture, so that the mesenchymal stem cells can be stably obtained, and the obtained mesenchymal stem cells have a bone differentiation tendency and lack the capability of differentiating to adipocytes. The mesenchymal cells obtained through in-vitro culture have application prospects in the field of cell therapy of bone related diseases.

The present invention provides a method for producing mesodermal lineage cells from an embryoid body, the method comprising: (1) a step for inducing an embryoid body into a differentiated embryoid body in which differentiation into a mesodermal cell lineage is promoted; and (2) a step for determining the solidity of the induced differentiated embryoid body.

The invention discloses a method for inducing directional differentiation of human induced pluripotent stem cells into mesenchymal stem cells in stages and application of the method, and develops a mesenchymal induction system which has definite components, is reasonably added with induced differentiation factors and has a reasonable concentration and is subjected to induced differentiation in stages. The induced pluripotent stem cells are regulated to efficiently and directionally induce and differentiate into the mesoderm cells, and then the mesoderm cells are directionally induced to induce and convert into the MSCs, so that the differentiation period of inducing and differentiating the human induced pluripotent stem cells into the mesenchymal stem cells can be shortened to 2-3 weeks, the differentiation rate is high, and the batch consistency can be ensured.

This invention provides a method for differentiating SAN-like cells from pluripotent stem cells in vitro. The method for differentiating SAN-like cells from pluripotent stem cells in vitro includes S1, providing a population of pluripotent stem cells; S2, performing a first contact culture on the pluripotent stem cell population to obtain mesodermal cells; S3, performing a second contact culture on the mesodermal cells to obtain pSHF cells; and S4, performing a third contact culture on the pSHF cells to obtain SAN-like cells. This invention solves at least one of the following problems in the prior art: the induction efficiency of sinoatrial node (SAN)-like cells from pluripotent stem cells is generally low, usually only 30%–50%; and the differentiation cycle is long, the operation steps are cumbersome, and a variety of small molecule additives are required. In addition, the differentiation products contain a large number of NKX2.5 positive atrial or ventricular myocytes, resulting in insufficient purity, which still requires further cell separation and purification.

To enable the construction of specific disease models involving multiple cell types and the development of screening systems for specific diseases, we provide a method for producing organoids containing ectoderm and mesoderm cells. [Solution] A method for producing organoids containing ectoderm cells and mesoderm cells, comprising the step (1) of culturing a cell aggregate containing pluripotent stem cells in suspension for at least two days in a culture medium containing a ROCK inhibitor and a TGFβ inhibitor.