29 results about "Developmental biology" patented technology

The invention discloses a preparation method of a mouse skin organoid, which comprises the following steps: separating dermis and epidermis of an E18.5 mouse to obtain dermis cells, mixing the dermis cells with back skin cells of an E14.5 embryo mouse, mixing by using Matrigel to form cell spheres, and gradually forming the skin organoid in in-vitro culture; the organoid is mainly applied to batch production of organoid containing hair follicles. The invention relates to the technical field of stem cells and regenerative medicine. According to the preparation method of the mouse skin organoid, a cell source is improved in the experiment process, the mouse skin organoid containing hair follicles can still be formed after cells in the early development stage are added, and related developmental biology research is carried out. And the source of another dermal cell is not limited to late pregnancy, and also can be a newborn mouse, so that the source of seed cells is expanded, and the cost is reduced.

The invention provides an antheraea pernyi fresh-keeping method which comprises the following steps: S1, dividing a first group and a second group with different freezing response characteristics according to development biological characteristics of antheraea pernyi; the freezing response characteristic is represented by at least one parameter of tissue moisture content, methylene blue penetration depth and body wall elasticity modulus; s2, differential preprocessing is applied to the first group and the second group, so that the difference of the first group and the second group on any characterization parameter does not exceed a preset threshold value; s3, combining the two groups of tussah and freezing; in the first cooling stage, cooling from 0-1 DEG C to-10 DEG C at the speed of 0.8-1.2 DEG C / min; in the second cooling stage, cooling is conducted from-10 DEG C to-30 DEG C at the speed of 0.45-0.55 DEG C / min; in the third cooling stage, cooling from-30 DEG C to-45 DEG C at the speed of 1.8-2 DEG C / min; and S4, after freezing is completed, vacuum packaging is performed, and storage is performed at minus 45-minus 40 DEG C. According to the fresh-keeping method for the tussah, the integrity of a cell structure is maintained, juice loss is reduced, and abnormal release of endogenous enzyme is inhibited, so that nutritional ingredients are better reserved in the long-term freezing storage process, and the sensory characteristics of compact meat, uniform color and luster and no peculiar smell are maintained.

The application discloses a single-cell transcriptome data pseudo-time trajectory analysis method and a computer system, which comprises the following steps: 1) calculating a gene explicit comparison advantage matrix; 2) obtaining a gene similarity matrix by similarity and constructing a gene network; 3) taking an initial node in the gene network, starting random walking from the initial node, recording each gene walked through to form a gene text composed of gene sequences; 4) converting the gene text into a gene word vector; 5) adding all single-cell expressed gene vectors with expression as weight to form a sum vector as a word vector representation of the single cell in the gene space; and 6) visualizing all cell vector representations to obtain an embryo cell development pseudo-time trajectory result. The application provides an analysis basis for identifying different rare cell subtypes in tissues and variant genes of different cell subtypes and has a wide and important application prospect in the fields of tumors, developmental biology and life science.

The application discloses a multi-agent based spaceomics tissue dynamics reconstruction and simulation perturbation method, and belongs to the technical field of computational biology and bioinformatics. The method models each cell as an agent, constructs continuous dynamic trajectories from the starting slice to the ending slice through unbalanced optimal transport and neural ordinary differential equations, uses a multi-agent reinforcement learning framework to make each agent decide the fate of proliferation, apoptosis, differentiation or migration according to the local microenvironment state, and combines a reward function to optimize the generation of missing intermediate tissue states; then, the cosine similarity is used to quantify the alignment degree of ligand-receptor communication signals and cell movement to identify key regulatory pairs, support computer simulation perturbation experiments on genes, communication pathways or cell populations. The application can reconstruct three-dimensional / four-dimensional tissue dynamics with high fidelity from sparse slices, analyze microenvironment regulation mechanisms and predict perturbation effects, and is suitable for spaceomics data analysis in the fields of developmental biology, oncology and the like.

This invention relates to a medaka hatching enzyme, its preparation method, and its application. The preparation method of the medaka hatching enzyme includes the following steps: 1) collecting fertilized medaka eggs; 2) incubating the fertilized medaka eggs with embryo culture medium in a 26℃ incubator for 6 days, and collecting mature embryos; 3) adding NaCl solution and Tris-HCl buffer (pH=8.0) to the mature embryos, grinding them, collecting the filtrate, and allowing it to stand overnight at 4℃; 4) centrifuging the filtrate at 15000 rpm for 15 minutes, and extracting the supernatant to obtain the medaka hatching enzyme solution. The preparation method is simple to operate, and the obtained medaka hatching enzyme has high activity, which can help to quickly remove the medaka eggshell artificially. The naked medaka embryos obtained by this method can be directly used for in vivo imaging and in-situ quantitative analysis, and embryo model construction, providing support for subsequent genetic, developmental biological, and toxicological studies of medaka embryos.

This invention provides an application and method of lysophosphatidylcholine (LPC) in promoting early development of reconstructed embryos, belonging to the field of embryo engineering technology. The invention first prepares reconstructed embryos, then adds lysophosphatidylcholine to the culture system for culturing. Lysophosphatidylcholine effectively promotes early development of the reconstructed embryos; at an optimal concentration of 5 μM, the blastocyst development rate increases to 24.56%, providing a novel and operable biochemical intervention target to solve the problem of low reconstructed embryo culture efficiency. This invention not only fills the gap in our understanding of the mechanism of action of LPC in this field, but also achieves controllability and reproducibility of the technical solution through precise definition of the optimal concentration. It provides key parameters for establishing standardized culture procedures and provides a more solid foundation for downstream applications such as developmental biology research and disease model construction that rely on high-quality reconstructed embryos, significantly accelerating the process of related scientific discovery and application development.

The invention relates to the technical field of crop development biology, in particular to a method for improving the germination speed and uniformity of wheat seeds indoors. In order to improve the condition of experimental errors caused by slow germination rate and inconsistent uniformity of wheat seeds, the germination speed and uniformity of the wheat seeds are remarkably improved through a method of cutting and damaging seed coats by virtue of endosperm part and soaking in ddH2O warm water, and the method is suitable for different types of wheat seeds, is low in cost and extremely simple to operate, and is suitable for popularization and application. The method is easy to master, popularize and apply, and is a universal and efficient new method.

This invention discloses a composition for the differentiation of mesenchymal stem cells (ADSCs) into keratinocytes and its application. The composition comprises: a Wnt signaling pathway activator, a TGF-β signaling pathway inhibitor, a bone morphogenetic protein pathway activator, and at least one of retinoic acid and a retinoic acid agonist. The small molecule composition provided by this invention is meticulously designed to gradually guide the fate transition of ADSCs by mimicking key signaling events during embryonic development. This strategy is not a simple compound screening, but a rational design based on a deep understanding of developmental biology, achieving precise regulation of cell lineages in vitro by reproducing the natural developmental process. The composition of this invention is not a simple superposition of components, but rather achieves cell fate transition by precisely regulating the balance of signaling pathway networks.

The invention belongs to the technical field of molecular biology, and particularly relates to a self-guiding type stpegRNA, a recombinant plasmid containing the self-guiding type stpegRNA and a CRISPR editing method for cell lineage tracking. The core of the invention is a DNA sequence, after the DNA sequence is transcribed into pegRNA, editing and self-extension of the DNA can be re-identified and mediated, and the sequence is characterized in that 5 '-3' sequences at the head end and the tail end of the pegRNA are basically consistent, and a pegRNA skeleton is subjected to homing modification. The development of the Infinite Printer marks the new era of the CRISPR pedigree tracking technology from limited recording to infinite analysis. Through fundamental reconstruction of an editing mode, collaborative breakthrough of capacity, resolution and safety is realized, and a brand new normal form is provided for research of developmental biology, oncology and regenerative medicine. In the future, along with the improvement of editing efficiency and the innovation of a calculation algorithm, the system is expected to become a standard tool for analyzing a complex biological system, and plays a reform role in the fields of personalized medical treatment, early diagnosis of diseases and the like.

The invention discloses a microglial cell and neurosphere co-culture system as well as a construction method and application thereof, and belongs to the technical field of cell biology and developmental biology. According to the invention, after the microglial cells and the neurospheres are mixed, three-dimensional space mutual embedding of the microglial cells and the neurospheres is realized through a gravity settling method, and stable co-culture of the microglial cells and the neurospheres is realized, so that the physiological environment in the brain and the natural interaction of neurons-immune cells in the brain are simulated more truly. The construction method of the microglia and neurosphere co-culture system is simple to operate, does not need to depend on a complex three-dimensional culture system or a specific scaffold material, and realizes construction of the stable and extremely-strong-repeatability co-culture system of the microglia and the neurosphere; the method is of great significance to deep study of physiological and pathological mechanisms of the nervous system.

The application discloses a kind of based on joint adjacency matrix spatial transcriptome data clustering method, belong to computer science and bioinformatics field.The method constructs a fusion SPOT-SPOT space adjacency matrix, SPOT-GENE expression matrix and GENE-GENE gene co-expression network matrix, to realize the adaptive alignment of multi-modal data in structure.On this basis, by introducing the graph auto-encoder with residual connection and multi-task learning mechanism, joint learning node low-dimensional embedding and model parameters, finally realize the accurate identification of spatial functional domain.The application constructs a complete technical process from multi-source data integration, graph structure modeling, feature learning to spatial domain identification, can accurately analyze the spatial organization mode and functional state of cell in tissue sample, provides a powerful computing tool for developmental biology, tumor microenvironment and other researches.

The invention belongs to the technical field of organoid culture, and discloses a hydrogel-based bionic brain micro-tissue as well as a construction method and application thereof. The hydrogel-based bionic brain micro-tissue is prepared by controlling the proportion of methylacryloylated gelatin to methylacryloylated hyaluronic acid, the proportion of a photoinitiator, the proportion of graphene oxide and key parameters of photocuring treatment. The hydrogel-based bionic brain micro-tissue has the advantages of definite components, controllable structure and form, low cost, easiness in standardization and the like, and meanwhile, the hydrogel-based bionic brain micro-tissue shows the biological properties of high cell activity, excellent biocompatibility, good safety and outstanding physiological-like functions. The method can be applied to the fields of brain-like tissue construction, nervous system disease modeling and mechanism research, neurodrug pharmacodynamics research, drug neurotoxicity evaluation or developmental biology research and the like.

The invention discloses a high-sensitivity space transcriptome sequencing platform, and belongs to the technical field of space omics and biological detection. The method comprises the following steps: sequentially delivering two groups of orthogonal space tag polynucleotides on the surface of a fixed tissue slice by utilizing an improved polydimethylsiloxane micro-fluidic chip, realizing space labeling of nucleic acid through reverse transcription reaction, and performing splitting decomposition, purification, amplification and library construction, and then performing next-generation sequencing to obtain high-resolution space transcriptome data. The micro-fluidic chip comprises channel arrays which are perpendicular to one another, and two-dimensional barcode marking of three samples can be completed on a single glass slide at the same time. The method is reasonable in structural design, uniform in space label delivery and low in signal pollution, can remarkably improve the capture efficiency and the space positioning precision, realizes multi-sample parallel high-throughput space transcriptome sequencing, and is suitable for the fields of developmental biology, pathological tissue analysis, clinical molecular diagnosis and the like.

The application discloses a set of gRNA, an RNP complex, a construction method of a gene knockout zebra fish model and application thereof, and belongs to the technical field of genetic engineering. ccny gene knockout, ccnyl1 gene knockout, ccny / ccnyl1 double gene knockout zebra fish models, which provide an animal experiment basis for cyclin-related research, provide a good animal model for fully understanding cyclin-related disease research and treatment strategy exploration, and help researchers to deeply understand cyclin-related molecular mechanisms and cell processes, ccny and ccnyl1 have a good application prospect in developmental biology.

This invention discloses a spatiotemporal tissue dynamics modeling method based on graph neural networks and imbalanced optimal transport. Belonging to the field of bioinformatics, this invention acquires spatial transcriptome data from multiple time points, constructs a low-dimensional latent space for gene expression, and aligns the spatial coordinates. It models the continuous evolution of gene expression states using graph neural differential equations; calculates cell growth rate using imbalanced optimal transport to obtain information on cell number changes; and constructs a spatiotemporal vector field integrating gene expression, cell number, and spatial coordinates to predict tissue state at target time points. This invention achieves collaborative and continuous modeling of cell state, spatial location, and population size during tissue development and regeneration, enabling generative prediction of tissue conformations at future unobserved time points. Furthermore, it is robust to spatial transcriptome data at different resolutions, providing an efficient and reliable computational analysis tool for developmental biology and regenerative medicine research.

The invention provides a gene ucp1 related to cold resistance of fishes and application of the gene ucp1 in cold-resistant breeding of the fishes, and belongs to the technical field of gene engineering. Based on zebrafish developmental biology and molecular biology research, a gene ucp1 related to fish cold resistance is found, based on a CRISPR / Cas9 technology, a CRISPR / Cas9 gene editing target site of the ucp1 gene is constructed, a zebrafish embryo is converted, and transgenic zebrafish with ucp1 knocked out is obtained. Results show that knockout expression of the ucp1 can improve the cold resistance of the zebra fish and relieve fish fever reaction caused by viruses, which indicates that the ucp1 gene is a gene for regulating and controlling the cold resistance of the fish and has a wide application prospect in the aspect of breeding of the cold-resistant fish.

The invention discloses a noninvasive microscopic imaging system for dynamically monitoring embryonic development, and relates to the technical field of biomedical imaging. The system comprises an imaging module, a dynamic embryo monitoring module and an intelligent development analysis module, the imaging module adopts a near-infrared pulse and low-power visible light dual-light-source system, supports phase difference, dark field and fluorescence multi-mode imaging, and remarkably reduces phototoxicity; the embryo dynamic monitoring module maintains the stability of an embryo culture environment in real time through an integrated culture cabin and a high-precision environment control system; the development intelligent analysis module realizes automatic image segmentation and feature extraction based on an improved U-Net network, generates a control instruction in real time through embryo respiration feedback, contour tracking and an automatic focusing algorithm, and realizes undisturbed, continuous and intelligent monitoring of the development process. The method solves the problems of large light damage, strong environmental disturbance, dependence on manual interpretation and the like in the prior art, and is suitable for the fields of assisted reproduction, developmental biology, drug screening and the like.

The invention relates to the technical fields of molecular biology, developmental biology, cytobiology and medicine, in particular to application of a PPARG gene or / and an expression product thereof in preparation of a medicine for inhibiting hepatocellular carcinoma, a method and the medicine. According to the present invention, the non-nerve cell differentiation-promoting factor PPARG gene or / and the non-nerve cell differentiation-promoting factor PPARG protein is / are adopted to inhibit tumorigenicity and malignant characteristics of liver cancer cells; the invention provides a protein PPARG capable of promoting muscle fiber differentiation, which can promote cancer cells to differentiate into muscle cells, so that the cancer cells lose nerve stemness and further lose tumorigenicity, thereby inhibiting the development process of cancers and treating the cancers. The invention is a novel cancer treatment method developed based on the discovery of the inventor on the essential characteristics of cancer cells, namely the brand-new regularity of the nerve stemness.

The invention relates to the field of environmental toxicology and developmental biology, in particular to a reproductive development toxicity evaluation method based on plastic greenhouse pesticide combined contamination. The combined action toxicity of beta-cypermethrin and emamectin benzoate is systematically evaluated from multiple dimensions of hormone level, ovarian tissue structure, placenta function, progeny development, cell viability and mitochondrial state through the design of combining an in-vivo model and in-vitro cell verification, and the potential hazard mechanism of beta-cypermethrin and emamectin benzoate in the reproductive development process is disclosed. Therefore, a scientific basis is provided for pesticide toxicology risk assessment and safety limit value formulation.

The present disclosure generally relates to systems and methods for spatial barcoding or identification of molecules in a sample. Certain embodiments relate to multi-dimensional spatiomics tools with targeted or non-targeted biomolecules, highly parallel barcoding, and subcellular resolution. Some aspects may use light to introduce spatial barcodes into biomolecules and sequence them to decode their content and location within cells and tissue blocks. Spatial barcoding may be accomplished using a light beam that may address pixels in 3D space without any dead zone between the pixels. Characteristics of these non-targeted RNA measurements, tissue 3D analysis, parallel barcoding, no dead zone and the likelihood of single cell sequencing provide a new way to acquire molecular and spatial information from tissue, e.g., for discovery in cancer biology, immunology, neuroscience and developmental biology.

This application discloses a nuclear localization signal AoNLS and its application in enhancing CRISPR / Cas9 editing in zebrafish. The nuclear localization signal AoNLS is derived from the zebrafish proteome. Through bioinformatics mining and functional screening, a structurally unique amino acid, KRKREAE, was identified, with its amino acid sequence shown in SEQ ID NO:1. This sequence is composed of both basic and acidic amino acids, and its mode of action differs from the single charge-driven mechanism of traditional NLS, achieving a breakthrough improvement in nuclear input efficiency. The Pcs2-AoNLS-GFP system, constructed by fusing AoNLS with GFP protein, showed a 1.56-fold increase in nuclear input efficiency in zebrafish embryos compared to the traditional SV40 NLS. Simultaneously, the Pcs2-AoNLS-Cas9 system, constructed by fusing AoNLS with Cas9 protein, significantly enhanced the editing efficiency of the Stat3 and Amh target genes by 1.72 times and 1.82 times, respectively, compared to the Pcs2-SV40 NLS-Cas9 gene editing system. The AoNLS sequence provided in this application is short and highly efficient, and can be used as a core component of the next-generation gene editing tool nuclear delivery system, with good application prospects in the fields of aquaculture breeding and developmental biology research.

The invention discloses a culture method of a bone tissue organ, and belongs to the technical field of tissue engineering and regenerative medicine. According to the method, a spleen is used as a tissue engineering scaffold platform with complete blood circulation and an original microenvironment, the spleen is shifted to a subcutaneous position which is easy to operate, and a bone tissue donor (including small-particle-size tissue particles or large-volume bone end segments) with development potential is implanted into spleen parenchyma; the functions of blood supply, nutrition delivery and waste removal of the spleen are utilized to realize in-vivo macroscopic development and structural reconstruction of bone tissue organs. According to the method, the natural development process of bone tissues can be simulated, long-term survival and functional maturation of organoids are supported, and an efficient and reliable experimental platform is provided for drug screening, disease modeling, developmental biology research and transplanted donor preparation.

The invention discloses a novel application of palmitoleic acid and a culture solution for promoting in-vitro development and maturation of porcine oocytes, and belongs to the field of reproductive biology in developmental biology. The method can be used for preparing products for promoting in-vitro development and maturation of oocytes. According to the present invention, pig cumulus-oocyte complexes (COCs) are cultured under the in vitro condition, and the palmitoleic acid is added to the culture solution to explore the influence of the palmitoleic acid on the first polar body discharge rate, the cortical particle distribution, the mitochondrial distribution and the oocyte reactive oxygen species (ROS) level of the oocytes, such that the results show that the in vitro addition of the palmitoleic acid can improve the oocyte quality, and the in vitro addition of the palmitoleic acid can provide the research on the oocyte first polar body discharge rate, the cortical particle distribution, the mitochondrial distribution and the oocyte reactive oxygen species (ROS) level; and a good effect is achieved.

The invention relates to the technical fields of molecular biology, developmental biology, cytobiology and medicine, in particular to application of an MYOD1 gene or / and an expression product thereof in preparation of a medicine for inhibiting hepatocellular carcinoma, a method and the medicine. According to the invention, the non-nerve cell differentiation-promoting factor MYOD1 gene or / and protein is / are used for inhibiting tumorigenicity and malignant characteristics of liver cancer cells. The invention provides a protein MYOD1 capable of promoting muscle fiber differentiation, which can promote cancer cells to differentiate into muscle cells, so that the cancer cells lose nerve stemness and further lose tumorigenicity, thereby inhibiting the development process of cancers and treating the cancers. The invention is a novel cancer treatment method developed based on the discovery of the inventor on the essential characteristics of cancer cells, namely the brand-new regularity of the nerve stemness.

The invention provides a novel dynamic bar code technology for tracking a cell development tree and application thereof. According to the technical scheme provided by the invention, the cells can be dynamically provided with new random tags along with cell proliferation, meanwhile, the tags same as those of previous cells are reserved, the genetic relationship between the cells is effectively recorded, and the cell genealogy is truly constructed. The invention also discloses a tool capable of simultaneously marking proliferative cells and dynamically tagging new random DNA tags on the cells depending on cell proliferation, and a genetic tool capable of being induced to express fluorescent proteins and bar code sequences (and guide RNA). The bar code genetic tool capable of being used for revealing the in-vivo cell lineage development tree is constructed for the first time, and a brand new technical method is provided for research in the fields of developmental biology, regenerative medicine, oncology and the like.

The invention discloses a group of gRNA and RNP compounds, a construction method of a gene knockout zebrafish model and application of the gene knockout zebrafish model, and belongs to the technical field of gene engineering. The ccny gene knockout zebrafish model, the ccnyl1 gene knockout zebrafish model and the ccny1 / ccnyl1 double-gene knockout zebrafish model are respectively constructed, an animal experiment basis is provided for cyclin related research, a good animal model is provided for fully understanding cyclin related disease research and treatment strategy exploration, and the method has the advantages that the ccny gene knockout zebrafish model and the ccnyl1 / ccnyl1 / ccnyl1 / ccnyl1 / ccnyl1 / ccnyl1 / Researchers are helped to deeply understand molecular mechanisms and cell processes related to ccny and ccnyl1, a theoretical basis is provided for developmental biology, and the method has a good application prospect.

This invention provides a spatial transcriptome multi-slice alignment and domain identification system based on geometric distance matching, belonging to the fields of artificial intelligence and bioinformatics. Technical solution: Based on a geometric distance matching algorithm and constructed using an MVC architecture, it includes two core functional modules: multi-slice data alignment and spatial domain identification, as well as three supporting modules: front-end interaction, back-end service, and data processing. By identifying common cell types across multiple slices, Euclidean distance is calculated after standardizing cell spatial coordinates to achieve nearest neighbor matching. A similarity threshold is then used to filter high-matching cell pairs to complete multi-slice alignment. Beneficial effects: The system is compatible with multi-platform data and mainstream formats, provides 2D / 3D visualization interaction functions and result export services, and solves the problems of complex models, high resource consumption, and high usage thresholds in existing technologies. It achieves efficient and accurate multi-slice data alignment and spatial domain identification, and is suitable for multiple research scenarios such as developmental biology and oncology.

The utility model provides an oocyte separator which is characterized in that the oocyte separator comprises an injection assembly which comprises a cylinder, a piston and a piston rod, one end of the cylinder is provided with a nipple part, and the nipple part is communicated with liquid of the cylinder so as to suck separated oocytes and / or push out liquid through the nipple part; the piston is arranged in the cylinder and is in airtight fit with the interior of the cylinder, and the piston rod is connected with the piston and is arranged in the cylinder in a sliding manner; the cutting assembly comprises a blade; wherein the injection assembly and the cutting assembly are detachably connected. According to the utility model, the thin blade cutting technology is introduced, so that the treatment efficiency is improved, the oocyte acquisition efficiency is improved, the operation difficulty and cost are reduced, and powerful technical support is provided for research in the fields of reproductive biology, genetics, developmental biology and the like.

The invention provides a multi-omics rare cell annotation method and system, computer equipment and a medium, and belongs to the technical field of biological information and computers. The method comprises the following steps: constructing a heterograph containing cell, gene and chromatin accessibility peak value nodes by integrating single cell RNA sequencing and ATAC sequencing data; a pre-fusion and graph fusion two-stage strategy is adopted, deep extraction and cross-modal fusion are carried out on the multi-omics features, and high-order node embedding is obtained; and further constructing reconstruction, clustering alignment and intra-class similarity multi-objective loss functions, and realizing accurate identification and annotation of rare cell types through joint optimization. By adopting the multi-omics rare cell annotation method and system, the computer equipment and the medium, the limitation of single-mode information is overcome, the capturing capability of low-abundance cell subgroups is improved, and the method has good generalization and biological interpretability and can be applied to the fields of tumor microenvironment analysis, immune response mechanism research, developmental biology and the like.