19 results about "Stem cell biology" patented technology

Provided is a method for comprehensive evaluation of the biological potency of mesenchymal stem cells, comprising: simulating in vitro a scenario where mesenchymal stem cells enter an inflammatory site in vivo, detecting expression levels of PD-L1, IL-6, HGF, and IDO1 in the mesenchymal stem cells, and detecting inhibition rates of proliferation of immune cells and secretion of inflammatory factor TNF-α, so as to evaluate the immunomodulatory function and tissue regeneration function of the mesenchymal stem cells.

The invention discloses a stem cell medicine for pancreas islet repair and application, and belongs to the technical field of stem cell biology, and the stem cell medicine for pancreas islet repair is prepared by compounding a carrier, pancreatic progenitor cells and pancreas islet-like cells in proportion. The problems that in the prior art, due to the fact that stem cells are unstable in differentiation efficiency, poor in synergistic effect and poor in biocompatibility, islet injury cannot be repaired, and diabetes cannot be treated are solved. According to the present invention, the dual mechanisms of endogenous beta cell repair and exogenous beta cell supplement are adopted, and the beta cell function recovery and the group structure reconstruction are synchronously achieved through the reasonable ratio, such that the in-vivo blood glucose of the mouse is significantly reduced, the beta cell number and the glucose tolerance dose of the mouse are increased, and the low immunogenicity and the high anti-inflammatory property are simultaneously provided. The compound can be used for treating type I and type II diabetes mellitus, improving the insulin secretion function and relieving local inflammatory response of pancreas by promoting pancreas islet beta cell proliferation, and has application potential in treatment of type I and type II diabetes mellitus and pancreas islet repair.

The invention discloses an immortalized porcine fibroblast fat progenitor cell line as well as a construction method and application thereof, and belongs to the technical field of stem cell biology and cell engineering. Pig muscle is taken as a source, primary porcine fibroblast fat progenitor cells are obtained through separation and screening, pTERT and pCDK4 genes are overexpressed at the same time through a genetic engineering method, the cell line can break through the Harike limit, immortalization is achieved, the cell line is transferred to 43 generations at present, the cell is in a healthy and full long shuttle-shaped state, and the progenitor cells can be used as progenitor cells of the porcine fibroblast fat progenitor cells of the porcine fibroblast fat progenitor cells of the porcine fibroblast fat progenitor cells. Long-term in-vitro culture still has adipogenic differentiation capacity and can be used for preparing products such as cell culture meat.

The application belongs to the field of stem cell biology and relates to a culture method of a 3D nerve immune organoid containing microglia cells, which comprises the following steps: S1, inducing induced pluripotent stem cells in ectoderm direction and mesoderm direction respectively to obtain nerve-like embryoid bodies and yolk sac-like embryoid bodies; S2, culturing the nerve-like embryoid bodies to make them differentiate into nerves to obtain nerve ring structures, re-digesting the nerve ring structures to obtain nerve progenitor cell single cells, and culturing the yolk sac-like embryoid bodies to make them generate macrophage progenitor cells; and S3, fusing the nerve progenitor cell single cells and the macrophage progenitor cells and continuing to culture to form the 3D nerve immune organoid containing microglia cells. The brain-like organoid of the application can realize the following functions: exploring the control of microglia cells on the proportion of progenitor cells in the development stage, the change of phagocytosis, the control on the number of mature neurons, the influence on cell proliferation and apoptosis, and simultaneously accepting external stimulation and making corresponding functional stress changes.

The invention discloses a method for detecting activity of human induced pluripotent stem cell derived myocardial cell telomerase, a reference gene, a primer and probe combination and application, and belongs to the technical field of stem cell biology and molecular detection. According to the method, stably expressed 18s rRNA is taken as a reference gene, total RNA at different differentiation stages of human induced pluripotent stem cell derived myocardial cells is extracted and is subjected to reverse transcription into cDNA, then a specific primer probe combination is adopted, the gene expression quantity of telomerase reverse transcriptase and 18s rRNA is detected through Taqman fluorescent quantitative PCR, and the activity of telomerase is analyzed and evaluated in a normalized manner in combination with a 2-delta delta CT method. The method solves the problem of detection deviation caused by instability of traditional reference genes, and is simple and convenient to operate, good in repeatability and reliable in result. The method can be used for evaluating the differentiation efficiency of the human induced pluripotent stem cell derived myocardial cells, the maturity of the myocardial cells, drug screening and heart disease model research, and has important application value.

The invention provides a method for comprehensively evaluating the biological efficacy of mesenchymal stem cells, which comprises the following steps of: simulating a scene that the mesenchymal stem cells enter an inflammatory site in a body in vitro, detecting the expression quantities of PD-L1, IL-6, HGF and IDO1 of the mesenchymal stem cells, and detecting the inhibition ratio of immune cell proliferation and secretion of inflammatory factor TNF-alpha, so as to evaluate the biological efficacy of the mesenchymal stem cells. And evaluating the immune regulation function and the tissue regeneration function of the mesenchymal stem cells.

This invention provides the application of T-5224 in the expansion and / or functional enhancement of human circulating hematopoietic stem / progenitor cells, relating to the fields of stem cell biology and regenerative medicine. It is the first time T-5224 has been applied to the expansion of cHSPCs, opening up a new pathway to improve stem cell culture efficacy by regulating downstream transcriptional hubs of inflammatory / stress signals. Accompanied by the upregulation of HOXB4 gene expression, it significantly improves LIN... ‑ CD45RA ‑ CD34 + CD38 low / ‑ The cell expansion factor, maintaining cell stemness and regeneration capacity, provides a new approach to solving the technical problem of the difficulty in rapidly expanding human circulating hematopoietic stem / progenitor cells in in vitro culture while maintaining cell stemness and regeneration potential.

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.

This application relates to stem cell biology and regenerative medicine. Disclosed herein are agents and methods for converting non-chondrocyte cells to chondrocytes or chondrogenic ceils, related cells, related compositions, related products, and related uses.

This invention provides the application of phloroglucinol in the expansion and / or functional enhancement of human circulating hematopoietic stem / progenitor cells, relating to the fields of stem cell biology and regenerative medicine. Phloroglucinol possesses dual inhibitory capabilities against both NF-κB and JNK / c-Jun, effectively reducing the protein level of phosphorylated NF-κB p65 (Ser536) in cultured cells, accompanied by upregulation of HOXB4 gene expression, and significantly improving LIN... ‑ CD45RA ‑ CD34 + CD38 low / ‑ The expansion fold of cHSPCs cells, maintaining cell stemness and regeneration capacity, provides a new approach to solving the technical problem of the difficulty in rapidly expanding human circulating hematopoietic stem / progenitor cells in in vitro culture while maintaining cell stemness and regeneration potential.

The invention belongs to the field of stem cell biology, and relates to a culture method of a 3D neural immune organoid containing microglial cells, which comprises the following steps: S1, respectively inducing induced pluripotent stem cells in an ectodermal direction and a mesodermal direction to obtain a neural embryoid and a yolk sac embryoid; s2, culturing the neural embryoid to perform neural differentiation to obtain a neural garland structure, and re-digesting the neural garland structure to obtain a neural progenitor cell single cell; culturing the yolk sac embryoid to enable the yolk sac embryoid to generate macrophage progenitor cells; and S3, fusing the neural progenitor cell single cell and the macrophage progenitor cell, and continuously culturing to form the 3D neural immune organoid containing the microglial cells. The brain-like organ provided by the invention has the functions of exploring the control of microglial cells on the progenitor cell proportion, the change of phagocytic function, the control on the number of mature neurons and the influence on cell proliferation and apoptosis in the development stage, and meanwhile, the brain-like organ can accept external stimulation and make stress change of corresponding functions.

The invention relates to the field of stem cell technology and gene detection, and discloses a mesenchymal stem cell biological efficacy evaluation method and system.The method includes the steps that in the living cell state, amplification curve forms serve as the mandatory premise for judging the effectiveness of the gene expression quantity ratio, cross-batch correction is carried out in combination with reference cells, and the biological efficacy of mesenchymal stem cells is evaluated; meanwhile, core evaluation indexes are adaptively switched according to the intrinsic expression stability of the cells, and strong coupling judgment is carried out on amplification process dynamic information reflecting the paracrine activity of the cells and gene expression quantitative information representing the potency potential of the cells in the same technical action; the problem that in the prior art, evaluation timeliness and functional correlation are mutually restricted is solved, and immediate accurate evaluation of the cell efficacy is achieved.

This invention provides the application of JSH-23 in the expansion and / or functional enhancement of human circulating hematopoietic stem / progenitor cells, relating to the fields of stem cell biology and regenerative medicine. The small molecule compound JSH-23 effectively reduces the protein level of phosphorylated NF-κB p65 (Ser536) in cultured cells, accompanied by upregulation of HOXB4 gene expression, and significantly improves LIN. ‑ CD45RA ‑ CD34 + CD38 low / ‑ The cell expansion factor, maintaining cell stemness and regeneration capacity, provides a new approach to solving the technical problem of the difficulty in rapidly expanding human circulating hematopoietic stem / progenitor cells in in vitro culture while maintaining cell stemness and regeneration potential.

The application provides applications of IWR-1-endo and A-83-01 in promoting expansion and enhancing functions of human circulating hematopoietic stem / progenitor cells, relates to the technical field of stem cell biology and regenerative medicine, and IWR-1-endo and / or A-83-01 can promote expansion of cHSPCs; compared with a single reagent, the combination of the two has a synergistic promoting effect, significantly promotes expansion of cHSPCs, and maintains cell stemness and regenerative capacity, thereby providing a new idea for solving the technical problems in the prior art that human circulating hematopoietic stem / progenitor cells are difficult to be rapidly expanded in ex vivo culture while maintaining cell stemness and regenerative potential.

Aspects of the present invention include methods and compositions related to the modulation of molecules regulating the regenerative potential of cells and tissues in the embryonic state and the loss thereof in later fetal and adult stages of development. Said methods and compositions have uses in research in stem cell biology and in increasing regenerative potential in fetal and adult tissues otherwise incapable of regeneration.

The invention relates to the field of stem cell biology and regenerative medicine, in particular to a method for reversing the senescence state of mesenchymal stem cells through combined application of mechanical force stimulation and alpha-ketoglutaric acid treatment. Through combined application of mechanical force stimulation and alpha-ketoglutaric acid, expression of senescence-related markers and inflammatory factors can be remarkably reduced, cell proliferation and metabolic activity can be enhanced, osteogenic differentiation potential can be improved, secretion and function improvement of exosomes can be promoted, and the treatment effect is remarkably superior to that of only one method. When the MSCs treated by the method are used for caudal vein injection of old mice, the motor function, the hair state and the tissue aging level can be remarkably improved, and a systematic anti-aging effect is shown. The invention provides a safe, controllable and convertible new strategy which is suitable for anti-aging treatment and tissue regeneration.

The invention discloses a universal type definitive endoderm culture medium for inducing pluripotent stem cells to differentiate into definitive endoderm cells and a culture method, and relates to the technical field of stem cell biology and regenerative medicine. The universal typing endoderm culture medium comprises a basic culture medium, polyvinyl alcohol, an animal-origin-free B-27 supplement, a TGF-beta activator, a Wnt signaling pathway activator, a JNK-JUN inhibitor, a PI3K / mTOR inhibitor, L-ascorbic acid and a ROCK inhibitor, wherein the basic culture medium is used for providing nutritional ingredients required by cell growth; the universal typified endoderm culture medium comprises a typified endoderm culture medium 1 and a typified endoderm culture medium 2, according to the culture method, the general type typing endoderm culture medium is constructed to obtain the typing endoderm cell balls, and the general type typing endoderm culture medium is definite in component and does not contain animal-derived components, so that the general type typing endoderm culture medium meets the requirement of a drug supervision institution on chemical component limitation of cell treatment products; high-density culture is carried out on the typified endoderm cells through suspension culture, so that the method is more suitable for production requirements.

Aspects of the present invention include methods and compositions related to the modulation of molecules regulating the regenerative potential of cells and tissues in the embryonic state and the loss thereof in later fetal and adult stages of development. The methods and compositions have uses in research in stem cell biology and in increasing regenerative potential in fetal and adult tissues otherwise incapable of regeneration.

This invention belongs to the interdisciplinary field of stem cell biology and ophthalmic treatment, and relates to a method for preparing exosomes from human umbilical cord mesenchymal stem cells (hUC-MSCs) induced by cobalt chloride and their application in the treatment of dry eye. The method for preparing exosomes from hUC-MSCs induced by cobalt chloride includes the following steps: S1: Obtaining human umbilical cord mesenchymal stem cells. S2: Under normoxic conditions, adding the hUC-MSCs obtained in step S1 to a culture medium containing cobalt chloride, followed by induction culture for 2-10 hours. S3: Performing ultracentrifugation on the cell culture medium obtained in step S2 to obtain hUC-MSC exosomes. This invention, by culturing hUC-MSCs under normoxic conditions with cobalt chloride, can quickly and efficiently increase the content of miRNA-21-5p and TGF-β in exosomes, without requiring hypoxia equipment, resulting in low cost, simple operation, and exosome activity fluctuation ≤10%, significantly improving preparation efficiency and stability. Drugs prepared from the exosomes obtained by this invention have excellent effects in treating dry eye and have promising application prospects.