41 results about "Ectoderm" patented technology

Methods for differentiating pluripotent stem cells to neuroectoderm in dynamic suspension culture using small molecule or protein inhibitors of TGFβ / Activin / Nodal signaling and BMP signaling are provided. Also provided are methoc and protocols for differentiating pluripotent stem cells such as human embryonic stem cells first to neuroectoderm, then further to glial progenitor cells, and further to oligodendrocyte progenitor cells (OPCs), and compositions obtained thereby. The methods of the present disclosure reproducibly produce neuroectoderm progenitor cells by day 7 of the differentiation process, glial progenitor cells by day 21 of the differentiation process and OPCs by day 42 of the differentiation process.

Methods for differentiating pluripotent stem cells to neuroectoderm in dynamic suspension culture using small molecule or protein inhibitors of TGFβ / Activin / Nodal signaling and BMP signaling are provided. Also provided are methot and protocols for differentiating pluripotent stem cells such as human embryonic stem cells first to neuroectoderm, then further to glial progenitor cells, and further to oligodendrocyte progenitor cells (OPCs), and compositions obtained thereby. The methods of the present disclosure reproducibly produce neuroectoderm progenitor cells by day 7 of the differentiation process, glial progenitor cells by day 21 of the differentiation process and OPCs by day 42 of the differentiation process.

The present invention discloses a method for culturing vitreoretinal organoids in vitro, comprising the following steps: (1) amplifying and culturing pluripotent stem cells to a cell confluence of 50% to 90%; (2) on day 0, digesting the stem cells with a digestion solution; adding a neurosphere culture medium for culturing; adding matrigel for culturing, and then adding a neurosphere culture medium for culturing to obtain a cell suspension; (3) adding a ROCK pathway inhibitor to the cell suspension, and culturing in the neurosphere culture medium for 3 to 5 days to obtain ectoderm neurospheres; (4) replacing the neurosphere culture medium with a vitreous culture medium I, adding TGFB1 protein, and placing the cell suspension on a shaker for 25 to 28 days; (5) replacing the vitreous culture medium I with a vitreous culture medium II, and culturing the cell suspension for 60 to 300 days to obtain vitreoretinal organoids. The vitreous organoids of the retina cultured in vitro according to the present invention can simulate the vitreous in vivo.

The invention discloses a method for generating haploid sperm-like cells through in-vitro differentiation of pluripotent stem cells and application of the haploid sperm-like cells. Human pluripotent stem cells are firstly differentiated into ectoderm-like cells, then the ectoderm-like cells are spontaneously gathered and converted into primordial germ cell-like cells under the induction of ectoderm induction factors, and then the primordial germ cell-like cells and testicular sertoli cells are subjected to mixed culture to form recombinant testicular tissues; the recombinant testis tissue enters meiosis under the induction of nutrition limitation and retinoic acid, then meiosis is completed under the stimulation of meiosis second-stage induction factors such as testosterone, follicle-stimulating hormone and pituitary extract, and interaction between primordial germ cell-like cells and testis supporting cells is enhanced by using small molecules SB431542, so that the meiosis of the testis is enhanced. And finally inducing to generate haploid sperm-like cells. According to the invention, a new tool can be provided for researching a molecular mechanism of male reproduction, and an important thought can be provided for research and transformation of asthenospermia, oligospermia, drug screening and the like.

To provide ectodermal mesenchymal stem cells and methods for producing the same.SOLUTION: A method for producing ectodermal mesenchymal stem cells includes the step of selectively collecting, from peripheral blood of a subject administered with a peptide composed of an amino acid sequence of SEQ ID NO: 1, either (a) PDGFRα-positive and CD34-positive cells or (b) PDGFRβ-positive and CD34-positive cells.SELECTED DRAWING: None

Systems and methods for assessing cytotoxicity of cell culture media and devices. Such systems and methods include genetically modified stem cells that have been modified to express a reporter when a gene associated with pluripotent and / or trophoderm development is activated in the genetically modified stem cells.

This invention relates to the field of bioengineering technology, and in particular to a method for simultaneously collecting the inner cell mass and trophectoderm of mammalian blastocysts. The method includes the following steps: (1) after removing the zona pellucida of the blastocyst, the blastocyst is cut into an inner cell mass and a trophectoderm portion, and the cut trophectoderm portion is transferred to a culture medium for collection; (2) the inner cell mass portion obtained in step (1) is sequentially placed in an antibody treatment solution and a complement treatment solution for incubation; (3) the inner cell mass portion treated in step (2) is pipetted to obtain a purified inner cell mass. This method achieves simultaneous and efficient separation of two key cell lines from the blastocyst, fundamentally changing the traditional method's "resource waste" model of only obtaining the inner cell mass while discarding the trophectoderm, and providing more comprehensive experimental materials for embryonic development, placental biology, and stem cell research.

This invention discloses a method for predicting the outcome of in vitro fertilization-embryo transfer (IVF-ET) based on static embryo images, used to distinguish between biochemical pregnancy and clinical pregnancy. It belongs to the field of medical artificial intelligence and assisted reproductive technology and is applicable to single embryo transfer (SET) and dual embryo transfer (DET) scenarios. The method includes: acquiring a static image of at least one embryo to be transferred; preprocessing and standardizing the embryo image, and then inputting it into a feature extraction model based on a convolutional neural network. The model uses DenseNet-169 as the backbone network and combines it with a Feature Pyramid Network (FPN) for multi-scale feature fusion. When the input is a dual embryo image, a Trophectoderm–Inner Cell Attention (TEIC-Attn) attention module is introduced to adaptively weight and fuse the features of the two embryos to characterize the contribution of different embryos to the final transfer outcome. When the input is a single embryo image, the prediction is completed based on the corresponding embryo features. The model ultimately outputs a probability prediction result of whether the IVF treatment outcome is a biochemical pregnancy or a clinical pregnancy. Furthermore, this invention combines a gradient-based class activation mapping method to generate interpretable visual heatmaps, highlighting key morphological regions closely related to embryo transfer outcomes, such as the inner cell mass and trophectoderm, thereby providing clinicians and patients with intuitive and reliable decision-making support. This invention enables unified prediction and interpretation of single and double embryo transfer outcomes without relying on time-series data, improving the accuracy and interpretability of in-vitro fertilization treatment decisions and demonstrating promising clinical application prospects.

The application discloses a culture solution for indirectly inducing human induced pluripotent stem cells into mesenchymal stem cells, which is composed of a neuroectoderm induction culture solution, a mesenchymal stem cell differentiation culture solution and a mesenchymal stem cell maintenance culture solution. The neuroectoderm induction culture solution is composed of a Neurobasal culture medium, a DMEM / F12 culture medium, an N2 additive, a B27 additive, a GSK3 inhibitor and a TGF-beta inhibitor. The application further discloses a method for indirectly inducing human induced pluripotent stem cells into mesenchymal stem cells, which comprises the following steps: taking the induced pluripotent stem cells, culturing the induced pluripotent stem cells in the neuroectoderm induction culture solution for 6 days, then culturing the induced pluripotent stem cells in the mesenchymal stem cell differentiation culture solution for 7 days, and then culturing the induced pluripotent stem cells in the mesenchymal stem cell maintenance culture solution for 3-5 generations. The method can direct the induced pluripotent stem cells to differentiate into mesenchymal stem cells in a short time, and greatly shortens the culture time.

Tissue-specific progenitor cells representing the endoderm, ectoderm, and mesoderm lineages are generated from pluripotent sources. Progenitor cell generation is accomplished by contacting pluripotent stem cells with tissue-specific exosomes, microRNAs, proteins, and peptides, obtained by stressing the tissue under in vitro conditions. In vitro-generated tissue organoids are used as a source of "differentiation factors" for generating individualized progenitor cells. For example, to generate lung progenitor cells, allogeneic stem cells are implanted into decellularized cadaveric lung tissue to form lung structures, which are then exposed to various cellular stressors and conditioned media to differentiate the pluripotent stem cells into progenitor cells. Extracellular matrix can be obtained from decellularized structures seeded with regenerating cells and used to generate organoids, which are then subjected to stress to produce differentiation factors.

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 present invention relates to a method for promoting wound healing in a subject suffering from Ectodermal dysplasia in need thereof comprising a step of administering subcutaneously, intradermally or topically to said subject a therapeutically effective amount of a compound which restores the activity of p63. Inventors have performed a primary culture of patient keratinocytes suffering from ectodermal dysplasias with two compounds which restore the activity of p63 (e.g. STIMA-1 and / or PRIMA-1Met). They have shown that there is an important differentiation of the keratinocytes of said patient compared to the cells not treated with these compounds. They observed that the activity of p63 mutated is restored, thus the proliferation and differentiation of keratinocytes from the patient are activated. Moreover, inventors have used PRIMA-1Met by topical application on a young patient suffering from ectodermal dysplasias and shown that said patient presents an improvement on her hand. Typically, severe skin erosions (on hands and feet) are healing when PRIMA-1Met is administered topically on the hand.

Apparatus and associated methods relate to inducing ectodermal stimulation by substantially continuous topical application of a therapeutic dose of vascular growth induction agent (VGIA) and moisturizing compound. The moisturizing compound may, for example promote skin health and / or penetration of the VGIA to target cells. The VGIA may, for example, promote vascularization, neurostimulation, and / or oxygenation. In an illustrative example, the VGIA may, for example, include minoxidil. The moisturizing compound may, for example, include lactic acid. In some implementations, the VGIA and moisturizing compound may be substantially continuously topically applied by a cream. In some implementations, the VGIA and moisturizing compound may be substantially continuously topically applied by a controlled-release application device. Various embodiments may advantageously induce reversal of disease processes at the ectodermal level in affected peripheral body portions (e.g., feet, legs, hands).

This application relates to a method, apparatus, computer equipment, storage medium, and program product for the quantitative evaluation of trophectoderm. The method includes: acquiring target parameters such as the geometric parameters of the closed region containing the trophectoderm in the embryo to be evaluated, the total number of cells included, the cell area of ​​each cell, and the total number of adjacent cell pairs; obtaining standardized quantitative indicators, density indicators, and homogeneity indicators of the trophectoderm based on the target parameters; and obtaining evaluation results by quantitatively evaluating the trophectoderm based on the standardized quantitative indicators, density indicators, and homogeneity indicators. This application provides a calculable, standardized, and comparable quantitative evaluation method for trophectoderm, improving the objectivity, consistency, and repeatability of quantitative evaluation of trophectoderm in blastocysts.

The disclosure relates to the field of generating blastocyst-like 3D structure of cells or models. In particular the disclosure relates to an in vitro method of producing a blastocyst-like 3D structure of cells having cells of embryonic lineage forming at least two distinguishable epiblast-like structures and cells of extraembryonic lineage forming at least two distinguishable hypoblast-like structures. The disclosure also relates to the blastocyst-like 3D structure of cells so obtained and to several uses of the same, in particular as model of monozygotic and monochorionic twinning. Methods of screening of candidate agents that modulate the mammal embryonic development are also disclosed.

The invention provides a construction method of an embryo-like model, which comprises the following steps: culturing primitive state stem cells in vitro or injecting the primitive state stem cells into an immunodeficient animal body for differentiation to obtain the embryo-like model. According to the method, in the process of preparing the embryo-like model, cell factors and small molecular pathway interference factors do not need to be added, gene editing is not needed, only a single starting cell type is adopted, and the method has the advantages of being simple and high in repeatability, facilitates large-scale preparation and application, and has wide application prospects. And the prepared embryo-like model has all embryo external cell lineages and all embryo internal cell lineages except for trophoderm, and can simulate the development process from an implanted embryo of a Carbonate-resistant group 3 stage to a Carbonate-resistant group 9 stage nerve embryo, so that the embryo-like model can be used for preparing various progenitor cell lineages, modeling diseases, screening drugs and carrying out scientific research.

The invention relates to an embryonic development potential deep learning prediction method based on blastocyst transcription sequencing, which can overcome the difficulty of DNA / RNA co-sequencing for blastocyst trophoderm trace cells, and additionally obtain transcriptome RNA-seq data on the basis of not influencing the existing PGT-A technology to obtain chromosome CNV through DNA sequencing. Differential expression genes (DEGs) are found, and the developmental potential of an embryo is predicted by analyzing the expression profiles of the genes. A blastocyst pregnancy outcome prediction model is established and serves as a method for screening embryonic development potential clinically, pregnancy development potential prediction is rapidly carried out, pregnancy time can be shortened, multiple pregnancy failures are reduced, clinical efficiency is improved, and important contribution is made to the field of reproductive medicine.

The invention provides an iPSC (induced pluripotent stem cell) differentiation efficiency improvement method and application, and the method realizes the extensive improvement of the induction efficiency of different 2D / 3D (two-dimensional / three-dimensional) derivatives of an endoderm source, an ectoderm source and a mesoderm source by combining screening type subculture and Rock inhibitor retention type differentiation initiation.

The invention discloses an induction method for directionally differentiating hESC-melanocytes based on a neural crest lineage, and belongs to the technical field of cell engineering and cytobiology. According to the induction method, a complex differentiation process of embryonic stem cells-ectoderm-neural ectoderm-neural crest cells is abandoned, and the neural crest cells are directly obtained from embryonic stem cells, so that the differentiation process is simplified; the differentiation efficiency and controllability of the neural crest cells are remarkably improved, the obtained melanocytes have higher tyrosinase activity and melanin synthesis capacity, and a new way is provided for treating leucoderma by adopting a melanocyte transplantation method.

Provided herein are compositions of anorectal transition zone stem cells, to treat endodermal, ectodermal or mesodermal disease. Also provided are cellular compositions of the anorectal transition zone stem cells, in a pharmaceutically acceptable carrier.

The invention discloses a human protogut-like in-vitro model based on embryonic cells, a construction method and application, and belongs to the technical field of protogut-like model culture. In order to solve the problem that the formation process of human original stripes cannot be simulated in vitro, a completely defined co-culture system is developed, embryonic stem cells and various extraembryonic cell types are cultured together, the regulation and control processes of in-vivo amniotic ectoderm, trophoblast and extraembryonic mesoderm are simulated, and the formation process of the human original stripes can be simulated in vitro. The invention also discloses the effect of different extraembryonic cells, which are not known before, in adjusting embryonic cells. Besides, by utilizing the advantages of the micro-engineering technology, the interaction between different extraembryonic cells and embryonic cells is reconstructed in space and molecule, and it is proved that the formation process of human original stripes can be summarized only through coordinated regulation and control of the extraembryonic cells. And a convenient in-vitro research platform is provided for analysis of human early-stage protointestine formation.

This method for producing mesenchymal cells of the ventral hindgut mesoderm comprises: culturing pluripotent stem cells in an induction medium A containing a TGF-β inhibitor, a Wnt agonist, and a fibroblast growth factor to induce differentiation into epiblast-like cells; culturing the epiblast-like cells in an induction medium B containing a Wnt agonist, a fibroblast growth factor, and retinoic acid to induce differentiation into caudal epiblast-like cells; culturing the caudal epiblast-like cells in an induction medium C containing a Wnt agonist, a bone morphogenetic protein, and a TGF-β family member to induce differentiation into posterior primitive streak cells; and culturing the posterior primitive streak cells in an induction medium D containing a Wnt agonist, a fibroblast growth factor, a bone morphogenetic protein, and a hedgehog signaling agonist to induce differentiation into mesenchymal cells of the ventral hindgut mesoderm.

The inventors discovered that ectodermal mesenchymal stem cells circulating in peripheral blood that are induced by necrotic tissue damage contribute to the regeneration of damaged tissue. On the basis of this discovery, provided are ectodermal mesenchymal stem cells, a method for producing the same, and a screening method for a substance having pluripotent stem cell inductive activity, the screening method using cells induced by necrotic tissue damage in peripheral blood as an index.

The present invention relates to: a method for differentiating stem cell-derived hepatocytes by using fasudil and gelatin; and the like. The method for differentiating stem cell-derived hepatocytes, according to the present invention, induces, by means of fasudil, the differentiation of stem cells into endoderm in an early stage of differentiation of stem cells into hepatocytes, inhibits lipid droplet formation through high-viscosity gelatin coating of a culture container, activates mitochondrial functions, up-regulates the expression of endoderm genes, and down-regulates the expression of ectoderm and mesoderm genes, thereby improving the efficiency of differentiation of stem cells into hepatoblasts. In addition, low-viscosity gelatin is added to the medium in a differentiation and maturation (late) stage so as to suppress the accumulation of lipid droplets, mitochondria are activated to improve the efficiency of differentiation of hepatoblasts into hepatocytes, and the function of differentiated hepatocytes is enhanced to improve the efficiency of differentiation of stem cells into hepatocytes. Therefore, hepatocytes obtained using the method are expected to be effectively used as a cell therapeutic agent, a transplant, and the like for treating liver diseases.

The present invention provides a method and application for improving the differentiation efficiency of iPSCs. This method combines screening-based subculture with Rock inhibitor-retaining differentiation initiation to achieve a comprehensive improvement in the induction efficiency of different 2D / 3D derivatives of endoderm, ectoderm, and mesoderm origin.

The present disclosure provides a method for obtaining RPE (retinal pigment epithelium) cells from iPSCs (induced pluripotent stem cells). The method involves: (a) generating embryoid bodies from a culture of iPSCs, in which the embryoid bodies are in non-adherent suspension culture, (b) plating the embryoid bodies on a culture dish coated with a suitable extracellular matrix in differentiation induction media (DIM), in which the DIM contains at least one WNT pathway inhibitor and at least two SMAD pathway inhibitors, (c) culturing the neuroectoderm lineage in differentiation propagation media (DPM) for rosette formation, (d) culturing the rosettes of step (c) in Retinal Pigment Epithelium Maturation Media (RPEMM) to facilitate retinal progenitor cells formation, and (e) plating the retinal progenitor cells of step (d) on a culture dish coated with suitable extracellular matrix in RPEMM to obtain RPE cells.

This invention discloses a BSH fluorescent probe PET-CT nine-square grid method for determining boron concentration in cancer cells. Using a BSH fluorescent probe PET-CT instrument, tumor cells from the endoderm, mesoderm, and ectoderm are selected according to the nine-square grid theory, and then their IC50 and PET-CT values ​​are measured separately. The significance of this invention is to assist in achieving precise treatment with BNCT capture therapy, maximizing the therapeutic effect of BNCT therapy, and avoiding the ineffective treatment caused by clinicians and dosimeters misselecting boron drugs, thus preventing patients from undergoing repeated radiotherapy, increasing suffering, and extending treatment time and costs.

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.