880results about "Germ cells" patented technology

The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1 and application thereof, and relates to the technical field of biology. The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1. The nucleotide sequence of the enhancer RNA molecule MZGAe1 is as shown in SEQ ID NO: 1; meanwhile, the invention further provides a specific sgRNA sequence for activating the molecule in a targeted manner and a recombinant vector of the specific sgRNA sequence. Expression of endogenous MZGAe1 of cells is specifically activated by adopting a CRISPR activation technology, and conversion of mouse embryonic stem cells to bicellular-like cells can be efficiently promoted. By providing a brand-new endogenous targeted MZGAe1 accurate activation tool, the efficient transformation of wild mouse embryonic stem cells to bicellular cells can be realized only under the condition of endogenous activation of single enhancer RNA molecule MZGAe1, the proportion is at least increased by 5%, the expression of a totipotent marker gene ZSCAN4 is activated, and the expression of the totipotent marker gene ZSCAN4 is promoted. And an efficient and specific brand-new research tool and scheme are provided for researching zygote genome activation, embryonic development early events and cell reprogramming.

The present invention relates to a modified stem cell and an ex vivo method for preparing defined mature cells from genetic programming of said modified stem cell which can be carried out in absence of certain growth factors, as well as a kit allowing to transform stem cells, and the different uses of the mature cells, for example for in vitro drug screening and toxicology or as cultivated meat. The genetic programming involves a molecular switch between two mutually exclusive programs of stem cell expansion and differentiation. The invention may be used for stem cells or other progenitors cells of any type, from any eukaryotic organism, but finds particular application in pluripotent stem cells from humans and livestock animals.

Disclosed herein are universal donor stem cells and related methods of their use and production. The universal donor stem cells disclosed herein are useful for overcoming the immune rejection in cell-based transplantation therapies. In certain embodiments, the universal donor stem cells disclosed herein do not express one or more MHC-I and MHC-II human leukocyte antigens. Similarly, in certain embodiments, the universal donor stem cells disclosed herein do not express one or more human leukocyte antigens (e.g., HLA-A, HLA-B and / or HLA-C) corresponding to MHC-I and MHC-II human leukocyte antigens, thereby rendering such cells hypoimmunogenic.

The invention provides a cryopreservation and resuscitation method for induced differentiation of pluripotent stem cells to metaphase cells and application, and the cryopreservation method comprises the following steps: (1) the initial cells are pluripotent stem cells, and are induced and differentiated to the metaphase; (2) recovering the cells by using a mild digestive enzyme, counting, and resuspending the cells in a cryopreservation solution; and (3) carrying out programmed cooling to-80 DEG C, and then transferring into liquid nitrogen for preservation for later use. According to the cryopreservation method and the cryopreservation liquid provided by the invention, the cryopreservation survival rate of the differentiated metaphase cells is obviously improved, apoptosis or irreversible stress injury induced by a traditional cryopreservation liquid is avoided, and the problems of large cell injury and low survival rate caused by using a general cryopreservation liquid in the prior art are obviously improved. The resuscitated cells keep good differentiation potential, can smoothly form a mature renal unit structure, and solves the problem of subsequent differentiation failure in the prior art.

The invention provides a method for overspeed preparation of gene edited goldfish. The method comprises the following steps: S1, obtaining gobiocypris rarus without primordial germ cells as receptor fish; s2, designing a target spot for the goldfish amh gene, introducing a Cas9 / gRNA complex into a goldfish fertilized egg, performing sequencing verification to obtain a mutant individual, and feeding the mutant individual as a donor fish; s3, taking gonads of donor fish, cutting into pieces, performing enzymolysis digestion, filtering, and performing density gradient centrifugation to enrich donor germline stem cells; s4, transplanting the germline stem cells of the donor fish into the receptor fish body, and feeding to obtain the gene edited goldfish sperms and ova. S5, hybridizing and feeding goldfish amh mutant sperms and ova produced by gobiocypris rarus to obtain gene edited goldfish individuals. According to the invention, gobiocypris gobiocypris is used as a receptor, the cross-subfamily gene edited goldfish is used as a donor, and an efficient, stable and easy-to-popularize method for preparing the gene edited goldfish is provided by utilizing a belly borrowing reproduction technology.

The present invention relates to the new use of an exosome secreted by hepatocyte lineage cells differentiated via the directed induction of embryonic stem cells, and in particular relates to the use of the exosome in the preparation of a drug for treating liver diseases. The exosome is secreted by hepatocyte lineage cells differentiated via the directed induction of embryonic stem cells, and the hepatocyte lineage cells are hepatic progenitor cells and / or mature hepatocyte-like cells.

The invention provides a method for evaluating drug enterotoxicity in vitro through an intestinal organ model, and relates to the field of biological medicine. Comprising the following steps: preparing six different culture media, culturing the hESC by using the different culture media, inducing the hESC to respectively perform directional endoderm cell differentiation, posterior intestinal bulb differentiation, intestinal organ differentiation and intestinal organ maturation, and evaluating the drug enterotoxicity by using a prepared intestinal organ model. The intestinal organ model prepared by the invention is similar to human intestinal cells in composition, can accurately reflect the toxic effect of the medicine in the physiological environment in the human body, can be repeatedly obtained in quantity, and avoids clinical risks and ethical constraints; the sensitivity of drugs with different enterotoxicity can be reflected through cell viability, and the method is suitable for rapid evaluation of in-vitro high-flux drug enterotoxicity.

The invention provides a serum-free stem cell culture medium and a preparation method thereof, and the culture medium realizes long-term stable culture of stem cells under a completely serum-free condition by optimizing basic culture medium components and adding a specific growth factor combination, an extracellular matrix simulant and a metabolism regulator. The culture medium comprises a basic culture medium, a recombinant human protein substitute, a cell adhesion promoting factor, a growth factor combination, an antioxidant and a metabolism regulator. Compared with the prior art, the invention has the following advantages: 1) animal-derived components are completely avoided, and the immunogenicity and the difference between batches are reduced; 2) maintaining the dryness of the stem cells through a specific growth factor combination; 3) adding a metabolism regulator to optimize cell energy metabolism; 4) the cost is obviously lower than that of commercially available like products; and 5) supporting long-term culture of stem cells without spontaneous differentiation. The invention also provides a preparation method of the culture medium and a method for culturing stem cells by using the culture medium.

The invention discloses a vector system and method capable of converting mouse embryonic stem cells into embryonic-like cells in a 2-cell period, the vector system comprises a mouse Zfp352 gene sequence, and the nucleotide sequence of the mouse Zfp352 gene is shown as SEQ ID No.1. The invention also discloses a method for converting the mouse embryonic stem cells into embryonic-like cells in a 2-cell period. The invention creatively discovers that through overexpression of the Zfp352 in the mouse embryonic stem cells, the efficiency of converting the mouse pluripotent embryonic stem cells into the totipotent embryonic-like cells in the 2-cell period can be remarkably improved.

The present application relates to a kind of RIC cells and its preparation method and application, especially it is related to the lineage-specific differentiation of pluripotent or multipotent stem cells, specifically related to a kind of RIC cell obtained from stem cell differentiation and its preparation method and application.

The invention discloses a bovine oocyte in-vitro maturation culture solution as well as a preparation method and application thereof, and belongs to the technical field of animal reproduction and embryo engineering. The preparation method of the bovine oocyte in-vitro maturation culture solution comprises the step of adding mitoquinine mesylate into a conventional IVM culture solution, wherein the adding concentration of the mitoquinine mesylate is preferably 50 nmol / L. The bovine oocyte in-vitro maturation culture solution provided by the invention can effectively reduce the oxidative stress reaction of the oocytes in the in-vitro culture process, enhance the mitochondrial function and remarkably improve the bovine oocyte in-vitro maturation quality and the embryonic development potential, so that the blastocyst rate is increased, and the problem that the development potential of the existing oocyte IVM culture solution is insufficient is solved.

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.

Systems for sorting sperm include a housing and a microfluidic system supported by the housing. The systems also include two or more inlets providing access to the microfluidic system to deliver sperm or semen and fluid to the microfluidic system, as well as an outlet for harvesting sorted sperm. The systems include a flow channel that provides a flow path for sperm from an inlet to an outlet while sperm travels against a fluid flow towards an outlet for harvesting. In the systems, fluid delivered to the microfluidic system via an inlet flows from the inlet towards one or more outlets, and the flow channel extending along the length of the microfluidic system provides a flow path for motile sperm to travel against the fluid flow towards a collection outlet for harvesting. In some embodiments of a system, the microfluidic system also includes a microporous filter arranged in the flow path between the inlet and the collection outlet to cause motile sperm traveling against the fluid flow to pass through the filter to reach the collection outlet for harvesting.

Disclosed are methods for producing controlled endometrial organoids using micropatterning techniques, and methods for using endometrial cells or organoids for diagnostic and predictive purposes, and for optimizing IVF and other infertility-related treatments, including embryo transfer and intrauterine insemination (IUI). These organoids grow in a spatially controlled manner and exhibit less heterogeneity than organoids grown directly in a three-dimensional culture. The specification also describes methods for predicting the success of embryo transfer or other IVF-related treatments, optimizing embryo transfer, improving embryo implantation in the endometrium, and performing embryo transfer in subjects. These methods include culturing endometrial organoids or endometrial epithelial cells, and contacting the endometrial organoids or cells with one or more blast-like cells. In some embodiments, the endometrial organoids or cells include endometrial cells obtained from subjects undergoing or considering IVF-related treatment.

This disclosure relates to cell therapy approaches for treating progranulin (PGRN) deficiencies with human induced pluripotent stem cell (hiPSC)-derived cells. Advantageously, the hiPSC-derived cells (e.g., microglia progenitor cells) described herein can cross-correct PGRN deficiencies in damaged or diseased cells while reducing the amount of endogenous cell ablation that is needed, as demonstrated by experimental results showing restoration of PGRN levels in GRN mutant cells and brain organoids.

Embodiments of the present invention may provide protecting, preserving, and even repairing biological cells during in vitro processing. A uniform environment such as a protective layer with exogeneous lipids may be formed around biological cells which may interact with the plasma membranes of the cells.

Provided is an isolated human naive pluripotent stem cell (PSC), wherein: (i) when the naive PSC is a female PSC, then said naive female PSC has two unmethylated alleles of an X-inactive specific transcript (XIST) gene; and (ii) when said naive PSC is a male PSC, then said naive male PSC has an unmethylated allele of said XIST gene. Also provided is a culture medium which comprises an ERK1 / 2 inhibitor, a GSK3beta inhibitor, a p38 inhibitor, a JNK inhibitor, a STAT3 activator and at least one agent selected from the group consisting of: bFGF, TGFbeta 1, a PKC inhibitor, a ROCK inhibitor and a NOTCH inhibitor; or at least agent selected from the group consisting of: a TGFR inhibitor, a FGFR inhibitor, a PKC inhibitor, a ROCK inhibitor and a NOTCH inhibitor.

The invention belongs to the field of stem cell culture, and relates to a pluripotent stem cell culture medium, which comprises an E6 basal culture medium, 2 ng / mL of TGF-beta1, 0-50 ng / mL of FGF-2, and 0.01-1 [mu] M of SUN11602. According to the technical scheme, proliferation and pluripotency of the hPSC are effectively maintained in a mode of combining small chemical molecules with low-concentration FGF-2.

To provide a composition useful for culturing and differentiating pluripotent stem cells.SOLUTION: A composition comprising cultured radial glial-like cells expressing at least one marker of Brain Lipid-Binding Proteins (BLBP), CD133 (Prominin 1), Abnormal Spindle Microcephaly Associated Proteins (ASPM), Baculovirus Inhibitors of Apoptosis Repeat Containing 5 (BIRC5 or Survivin), FAT atypical cadherin 1 (FAT1), Hes family bHLH transcription factors 5 (HES5), SRY-Box transcription factors 21 (SOX21), or PAX6 proteins, A composition wherein the radial-glial-like cells are or have been cryopreserved in a cryopreservation medium comprising Chroman1 and / or a derivative thereof, Emricasan and / or a derivative thereof, trans-ISRIB and a polyamine comprising putrescine, spermine and spermidine.SELECTED DRAWING: None

A sequential cell culture system and methods for the in vitro generation of human spermatozoa from differentiating spermatogonia (dSPGs) and spermatogonial stem cells (SSCs) is provided. The instant disclosure encompasses the discovery and optimization of distinct, stage-specific culture media and conditions that enable, for the first time, the complete progression of human spermatogenesis outside the body. The process includes: (1) inducing entry of dSPGs and SSCs into meiosis; (2) supporting meiotic progression and completion to round spermatids; and (3) promoting spermiogenesis and the formation of elongated, spermatozoa. The invention further provides compositions, systems, and protocols for each stage, as well as markers and methods for identifying successful progression through each developmental transition. This system enables, for the first time, the complete in vitro recapitulation of human spermatogenesis, providing a platform for research, infertility treatment, and assisted reproductive technologies.

The present invention relates to CD73 + CD44 + , CD90 + A method for generating a population of mesenchymal stromal cells (MSCs) is provided. + CD44 + , CD90 + MSCs are used in methods to generate terminally differentiated osteogenic, adipogenic, and chondrogenic cells from pluripotent stem cells (PSCs), which involve the use of a single agent, a WNT signaling pathway activator (e.g., a GSK3β inhibitor), in adherent cultures of PSCs.

This invention provides a method for propagating pluripotent stem cells (PSCs) by suspension culture in a bioreactor. [Solution] The method comprises the following steps: (i) adding a ROCK inhibitor (ROCKi) to pluripotent stem cells cultured in suspension in a bioreactor; (ii) adding a cell dissociation agent, thereby dissociating aggregates of pluripotent stem cells; (iii) diluting the cell dissociation agent added in step (ii) by adding a sufficient excess volume of culture medium to reduce the concentration of the cell dissociation agent to a concentration at which cell aggregates can be re-formed; and (iv) culturing the mixture obtained in step (iii) under appropriate conditions that allow for the proliferation of PSCs.

The present disclosure relates to methods, products, and compositions useful for deriving bovine induced blastoids. Methods are described that allow for juvenile stem cell aggregation and induced blastocyst formation. The invention further provides an aggregation culture medium and an induced blastocyst-like culture medium which are suitable for promoting aggregation of the young stem cells and preparing the induced blastocyst-like. The bovine induced blastoids can be used in selection and breeding procedures, including the replication of pre-implant embryos with desired characteristics and in vitro breeding including a cross-generation breeding cycle from parent germ cells and / or embryos to progeny embryos.

This disclosure features tissue storage, irrigation, and infusion media compositions, and methods of using such compositions, for example, the temporary storage of cells and tissues prior to implantation or re-implantation, or for wound irrigation, wound packing, surgical site irrigation, or intravenous infusion. The tissue storage, irrigation, and infusion media compositions disclosed herein are useful in a healthcare or veterinary setting.

Featured are methods and compositions for the in vitro maturation of oocytes. In particular, the disclosure features methods and compositions related to engineering a plurality of ovarian support cells (OSCs) for promoting the maturation of an oocyte in a medium. Such methods and compositions are particularly useful for assisted reproduction technology (ART) procedures.

The present invention provides a cardiac tissue model comprising cardiac tissue having at least one lumen or central cavity, wherein the cardiac tissue model comprises at least two different cardiac tissues selected from left ventricular tissue, right ventricular tissue, atrial tissue, outflow tract tissue, atrioventricular canal tissue, sinoatrial node tissue, and atrioventricular node tissue, the central cavity can be shared by at least two different cardiac tissues, and / or at least two different cardiac tissues comprise the ability to propagate calcium signaling connections and / or tissue contractions, a method of generating such a tissue model, and the use of the tissue model for screening purposes.

The invention discloses construction of a Guangdong small-ear spotted pig spermatogonial stem cell in-vitro culture system and screening of surface markers, and belongs to the technical field of cell culture.The technical scheme includes that testis are broken firstly, then enzyme digestion, density gradient centrifugal purification and differential adherent purification are conducted, and purified spermatogonial stem cells are obtained; inoculating the obtained spermatogonial stem cells to feeder layer cells and culturing in a culture medium; the culture medium is a DMEM culture medium added with FBS and growth factors; the mass fraction of the FBS in the DMEM culture medium is 8-12%, an enzyme digestion method, a density gradient centrifugation method and a differential adhesion method are combined for application, the Guangdong small-ear spotted pig spermatogonia stem cells with certain purity are obtained, cells enriched and purified in vitro can proliferate massively, and an in-vitro culture system of the Guangdong small-ear spotted pig spermatogonia stem cells is established.

The present disclosure provides methods of producing midbrain dopaminergic neurons (mDAs) and precursors thereof, mDAs and precursors thereof produced thereby, as well as compositions comprising these cells, and their use for the prevention and / or treatment of neurological diseases. The present disclosure also provides methods of isolating mDA and precursors thereof from cell populations using novel surface markers.

The invention discloses application of Kitl in promoting meiosis of germ cells cultured in vitro. By adding a small molecule Kit ligand (Kitl), germ cells (including primordial germ cells PGCs, primordial germ cell-like cells PGCLCs and the like) cultured in vitro are promoted to enter and complete the first meiosis earlier stage, meiosis key protein expression is improved, and homologous chromosome association and recombination efficiency is improved. The method has a wide transformation medical prospect in the fields of human assisted reproductive technology and reproductive medicine.

In a method for inducing differentiation into neuromuscular junctions according to the present disclosure, the neuromuscular junctions are induced by: preculturing human pluripotent stem cells in a medium containing activin, bFGF, a WNT signal inhibitor, a BMP signal inhibitor, and a ROCK signal inhibitor; then culturing the stem cells in a medium containing bFGF, a ROCK signal inhibitor, a WNT signal activator, and a TGFβ signal inhibitor to differentiate into neuromesodermal cells; culturing the neuromesodermal cells in a medium containing a BMP signal inhibitor, a TGFβ signal inhibitor, a retinoic acid activator, and an SHH signal activator to differentiate into skeletal muscle progenitor cells and spinal motor neuron progenitor cells in the same culture dish; and culturing these progenitor cells in a medium for cell maturation containing BDNF, GDNF, AA, and dbcAMP.