47 results about "Sexual differentiation" patented technology

The invention discloses a method used for inducing differentiation of human pluripotent stem cells into hepatocytes using a small molecular compound. The method comprises following steps: differentiation of human pluripotent stem cells into definitive endoderm cells is induced; differentiation of the definitive endoderm cells into hepatic progenitor cells is induced; and differentiation of the hepatic progenitor cells into mature hepatocytes is induced. Compared with the prior art, the method possesses following advantages: in the whole induced differentiation process, the small molecular compound, and culture mediums and additives with clear composition are adopted, using of cytokines and culture medium with unclear composition is avoided, the system is stable, and the repeatability is excellent; the cost of the small molecular compound is lower than that of cytokines, compared with conventional cytokine method, cost is reduced by 2 / 3, so that the whole induced differentiation processis lower in cost, and it is beneficial for further enlarged differentiation applications; the induced differentiation process takes 13 days, the time consumption is the shortest in whole small molecular compound induced differentiation schemes reported home and abroad; the concentration of adopted DMSO is low; and the cytotoxicity is low.

The invention aims to provide a method for sex induction of hermaphrodite shrimps. According to the method, a target gene in macrobrachium is silenced by using an RNA interference technology to realize sex induction of macrobrachium rosenbergii, wherein the nucleotide sequence of the target gene is SEQ ID NO:1. By virtue of the induction technology provided by the invention, a macrobrachium individual has sex reversion and becomes hermaphrodite macrobrachium rosenbergii, and the developments of a female reproductive system and a male reproductive system are realized in the body of the hermaphrodite macrobrachium rosenbergii, so that a set of effective macrobrachium rosenbergii sex induction technology is established. Meanwhile, by using the hermaphrodite shrimps cultured by the method provided by the invention, a good scientific research material and an animal sample can be provided for sex determination, sex differentiation and researches on reproduction and fertilization of shrimps and crabs.

This invention relates, e.g., to a method for differentiating mammalian (e.g., human) pluripotent stem cells (PSCs) into endothelial cells (ECs) in vitro, by plating a single-cell suspension of PSCs onto a suitable surface such as type IV collagen and culturing the cells with VEGF after which ECs can be harvested. A preferred embodiment of the method first cultures the cells without VEGF and then sequentially cultures the cells with VEGF. Differentiation can be enhanced by adding an inhibitor of transforming growth factor β to the culturing with VEGF.

The invention relates to an ex vivo method for obtaining a population of human melanocytes derived from human pluripotent stem cells comprising the step consisting of co-culturing human pluripotent stem cells with cells that support ectodermal differentiation in the presence of an agent that stimulates epidermal induction and an agent that stimulates terminal differentiation of keratinocytes. The invention also relates to human melanocytes obtainable by said method and to uses thereof in cell therapy and in screening assays.

The invention discloses a siRNA-IR sequence for sex reversal of male macrobrachium rosenbergii and application of the siRNA-IR sequence. Expression of an IR gene is inhibited by a siRNA interference technology to promote sex reversal of the male macrobrachium rosenbergii to obtain neo-females. IR gene fragments of macrobrachium rosenbergii are screened and obtained from macrobrachium rosenbergii transcriptome, and the siRNA is designed and synthesized according to a macrobrachium rosenbergii IR gene sequence, named siRNA-IR. The siRNA-IR is transformed into macrobrachium rosenbergii by a microscopic injection method before a sex differentiation stage to interfere and inhibit expression of the IR gene in the macrobrachium rosenbergii and promote sex interversal of the macrobrachium rosenbergii. Compared with a dsRNA interference technology, the siRNA technology has smaller injection dosage, and can reduce the cost, effectively reduce immunological stress reactions of post-larvae of themacrobrachium rosenbergii and increase the survival rate of neo-females.

The invention discloses an Eriocheir sinensis EsSXL gene, an amplification primer group thereof and an amplification method. A full-length cDNA (complementary Deoxyribose Nucleic Acid) sequence of the EsSXL gene is cloned from an Eriocheir sinensis testis tissue by using the primer group provided by the invention, and the EsSXL gene comprises two variable spliceosomes EsSXL1 and EsSXL2, and lays an important foundation for the research of the important function of the function of the Eriocheir sinensis EsSXL gene in a sex differentiation process of the Eriocheir sinensis EsSXL gene.

The invention belongs to the technical field of plant tissue culture, and relates to a method for establishing a Malus sieversii regeneration system based on callus differentiation. The method is completed through the steps of induction of calluses and adventitious buds, proliferation of the adventitious bud, rooting culture, and hardening and transplantation of seedlings. The method allows the callus induction rate to be 100%, the highest induction rate of the adventitious buds to be 30%, the rooting rate to reacj 85% and the survival rate of the seedlings to reach 90%. The method provides abasis for the germplasm preservation and genetic transformation study of Malus sieversii. The method is suitable for various environments, especially Xinjiang, Gansu and other places with low air humidity, greatly improves the survival rate of tissue culture plants, and saves time and labor.

An early-period gender judging method for soft-shell turtles relates to the field of gender differentiation and gender regulation of temperature-determining animals. The method comprises the following steps of weighing the soft-shell turtle which was hatched two weeks ago, soaking the soft-shell turtle in alcohol, acquiring and measuring external shape parameters, observing sexual glands of female and male turtles, determining gender of the soft-shell turtle through tissue slice observation, acquiring soft-shell turtle external characteristic parameters which are significantly related with the gender through statistical analysis, obtaining a regression equation and judging the gender of the newly born soft-shell turtle through the regression equation. Quantitative indexes for early-period gender judgment of the soft-shell turtle are presented. Gender judgment can be quickly and accurately performed through external data measurement without animal killing two weeks after hatching of the soft-shell turtle. The early-period gender judging method has advantages of high speed, high accuracy, no requirement for animal killing, and high operability.

The present invention encompasses methods, assays and kits for the diagnosis, screening and identification of Turner syndrome and other disorders of sexual differentiation in a human using single nucleotide polymorphisms present on the X and Y chromosomes.

The invention discloses a Macrobrachium nipponense MnIAG gene, and an amplimer group and an amplification method thereof. The full-length cDNA sequence of the MnIAG gene is cloned from the androgenic gland tissue of Macrobrachium nipponense through utilizing the primer group provided by the invention, and an important basis is established for the function of the Macrobrachium nipponense MnIAG gene, especially the important effect in sex differentiation.

The invention provides a Toona sinensis Roem regenerating system based on callus differentiation. The Toona sinensis Roem regenerating system based on the callus differentiation comprises the following steps: obtaining sterile explants of Toona sinensis Roem, putting the explants into an inducing medium, culturing to form calluses, culturing the calluses in a basal medium, then transplanting into a differentiating medium, culturing and subculturing to form buds, culturing the buds in the differentiating medium, subculturing in the basal medium after the buds grow more than 4-6 leaves, then transplanting into a rooting medium, rooting to form seedlings, culturing the seedlings in the rooting medium, hardening, transplanting and the like. The Toona sinensis Roem regenerating system based on the callus differentiation is high in propagating coefficient and high in propagating speed, and helps to promote cultivation of a fine variety; the inducing rate of the calluses is 100% and the survival rate of regenerated plantlets growing from the rooted seedlings is above 96%. The Toona sinensis Roem regenerating system based on the callus differentiation can be used for rapidly propagating a precious and fine Toona sinensis Roem variety, so that large-scale production of the fine variety is achieved, needs of shed cultivation or greenhouse cultivation are met, application of the fine Toona sinensis Roem variety is accelerated; therefore, the Toona sinensis Roem regenerating system based on the callus differentiation has a relatively high market value.

The invention provides methods and kits for monitoring the status of women's health, including HRT monitoring and determination or diagnosis of ovulation, contraception, pregnancy, menopause, polycystic ovarian disease, and female sexual dysfunction. The method comprises the steps of: (a) collecting a tear fluid from a female human; (b) determining the tear concentration of at least one hormone of relevance to female fertility, sexual differentiation or sexual dysfunction in a female human, wherein the tear concentration is diagnostics of the status of women's health.

The invention discloses a regulating agent capable of promoting the development and fruit setting of pistillate flowers, and a preparation method thereof. The regulating agent comprises 30 to 50% of calcium potassium nitrate, 15 to 25% of magnesium potassium nitrate, 0.1 to 1.0% of dihydrozeatin riboside, 0.1 to 0.5% of ethephon, 5 to 10% of boric acid, 1 to 2% of an emulsifier xanthan gum and 15to 50% of water. The regulating agent provided by the invention employs ethephon and dihydrozeatin riboside with promoting effects on the basis of the growth and nutrient demand characteristics of balsam pears and directed at problems occurred in the differentiation, development and fruit setting of pistillate flowers, so the regulating agent can change the sex differentiation direction of the flowers in an early development stage of the flowers and promote increase in the number of the pistillate flowers; meanwhile, the regulating agent supplements nutrients such as nitrogen, potassium, calcium, magnesium and boron needed for fruit swelling of the balsam pears so as to promote the rapid swelling of balsam pear fruit, and the purposes of eliminating abnormal balsam pears, increasing yieldand improving quality are achieved.

The invention relates to a method for simultaneously identifying the genetic sexes of litopenaeus vannamei at the DNA or RNA level. The early-stage genetic sex determination of prawns not only is of great significance in a production process, but also plays an important role in researching the molecular mechanisms of sex determination and sex differentiation. Previously reported prawn genetic identification methods can only use the DNAs of individuals to perform molecular identification, and no related method for identifying sexes by using DNAs and RNAs exists at present. According to the method for simultaneously identifying the genetic sexes of litopenaeus vannamei at the DNA or RNA level, three SNP markers with sex differences on sex-related genes are utilized, and the SNPs are typed, and therefore, the sexes of the individuals can be identified through the DNAs or RNAs of the individuals. With the method adopted, the obstacle that sex determination cannot be realized by using the RNAs of individuals in the past can be eliminated; a new sex determination method and means are provided; and a favorable tool is provided for subsequent sex determination and sex differentiation research.

The disclosure provides a method of generating a sterile sex-determined fish, crustacean, or mollusk. The method comprises breeding (i) a fertile homozygous mutated female fish, crustacean, or mollusk having at least a first mutation and a second mutation with (ii) a fertile homozygous mutated male fish, crustacean, or mollusk having at least the first mutation and the second mutation to produce the sterile sex-determined fish, crustacean, or mollusk. The first mutation disrupts one or more genes that specify sexual differentiation, the second mutation disrupts one or more genes that specify gamete function, and the fertility of the fertile homozygous female fish, crustacean, or mollusk and the fertile homozygous mutated male fish, crustacean, or mollusk has been rescued. The disclosure also provides methods of making broodstock for use in producing sterile sex-determined fish, crustacean, or mollusks, as well as the broodstock itself.

The invention discloses an immortalized human neural stem cell line and a preparation method thereof. The method comprises the following steps: (1) isolating and culturing primary human hippocampus-derived neural stem cells; (2) constructing a recombinant lentiviral vector containing L-myc gene; (3) carrying out packaging production of the above recombinant lentiviral and collecting supernatant; (4) selecting virus supernatant with appropriate concentration for transfection and screening of stem cells and constructing the immortalized human neural stem cell line. The preparation method of immortalized cell line provided by the invention is a novel stable and safe and effective immortalized coding strategy; the prepared immortalized human neural stem cell line has all the biological characteristics of primary neural stem cells, and also has the feature of normal three-line differentiation at the same time, can be successfully differentiated into neurons, astrocytes and oligodendrocytes,and the growth rate of such cell line and ability to form neurospheres are stronger than the growth rate of primary cells, and the cell line can still proliferate rapidly after 20 generations withouttumorigenicity.

According to the invention, prawns have obvious sex bimorphism; sex determination and differentiation of the prawns often occur in an early development stage; thus, sex determination and sex differentiation related genes need to be screened in the early development stage of male and female individuals. Since prawn individuals at the early development stage are extremely small, and cannot be distinguished in sex, so a screening method for male and female differential expression genes of early embryos or larvae of the prawns is not established yet. According to the invention, co-extracting of DNA and RNA of a single individual of a prawn zoae is realized; a sex probe is used for sex identification on a DNA level; RNAs of a same gender are mixed to respectively construct a male RNA library and a female RNA library in a zoae stage; a micro transcriptome sequencing technology is utilized to obtain the female and male larva differential expression genes; thus, the screening method of the male and female differential expression genes of the early embryos or larvae of the prawns is established. The screening method provided by the invention promotes the research on sex determination and sex differentiation of the prawns and provides a basis for analyzing a sex determination and differentiation mechanism.

The present invention relates to a VEGF-165 activator and use thereof for promoting stem cell differentiation. Alkaline phosphatase (ALP) is an early osteogenesis marker, and is mainly distributed incell membranes to promote cell calcification. The quantitative detection of the ALP can reflect the differentiation level of osteoblasts. If the activity of the ALP is higher, the higher activity of the ALP indicates that pre-osteoblasts have more obvious differentiation tendency to mature osteoblasts. The high expression of the activity of the ALP is an early marker of osteoblast differentiationand maturation. When the activity of the ALP is enhanced, bone formation is enhanced, and bone matrix mineralization is promoted. Therefore, the activity of the ALP is a good indicator for reflectingthe degree differentiation and the functional status of the osteoblasts. The osteogenic differentiation of hBMSCs can be promoted by activating BMP-2 protein expression in the hBMSCs by to-be-tested compounds 1 to 3. The osteogenic differentiation of the hBMSCs can be promoted by activating VEGF-165 protein expression in the hBMSCs by to-be-tested compounds 4 and 5.

The invention discloses a n establishment method of an all-male micropterus salmoides production system. The method comprises the following steps: (1) domesticating: domesticating the micropterus salmoides to eat artificial compound feed before sex differentiation of juvenile fish of the micropterus salmoides; (2) sex reversal: treating young micropterus salmoides by using hormone in a sex differentiation sensitive period, and reversing the young micropterus salmoides into physiological female fish with a reproductive function; (3) parent vitamin fortification: treating the parent fish with hormone, and adding vitamins into feed for nutrition fortification 3 months before sexual maturity of the parent fish; (4) test mating: mating physiological female fish and normal male fish for spawning, and deducing the sex genetic type of the hormone-treated parents according to the sex proportion of filial generations of the female fish and the normal male fish; (5) expanding of sex-reversal female fish: performing sex hormone treatment by using the same method, and when a test mating result is obtained, reserving a filial generation of the sex-reversal female fish as a breeding parent; and (6) taking individuals treated by sex hormones of the sex reversal offspring as a female parent group, taking normal male fishes as a male parent group, and establishing a production system capable of continuously producing all-male micropterus salmoides fries.

The invention relates to an early sex identification method for tortoises, belonging to the field of sex differentiation and sex adjustment and control of temperature-dependent sex determination animals. After being weighed, young tortoises, which are two weeks old, are immersed by ethyl alcohol; then, external morphological parameters are acquired and measured; male and female young tortoise gonads are observed; found from tissue section observation, the sexes are determined; external characteristic parameters of the tortoises obviously related to the sexes are obtained through statistical analysis; therefore, a regression equation is obtained; and the sexes of the newborn tortoises can be judged through the regression equation. Animals do not need to be slaughtered; sex identification of the young tortoises, which are about two weeks old, can be carried out through appearance; compared with a gonad tissue section method, the method is simple, high-efficiency and simple to operate; the accuracy rate is higher than 95%; the operability is high; tools required for performing sex identification are simple and easy to obtain; operation is simple; and the tools include a vernier caliper and a computer.

The invention provides a planting method for dwarfing and high-yield medicinal cannabis sativa. The planting method comprises the steps of preparation before field planting, field planting after seedling raising and management after field planting; management after field planting comprises a seedling growing period, a rapid growing period, synthesis promoting agent injection and a flowering period. Cannabis sativa seedlings are transplanted into a root control container for field planting, dwarfing and rational close planting of the cannabis sativa are achieved, the height of cannabis sativa plants is controlled to be 92-103 cm, the land space is saved, the photosynthetic area is effectively increased, the yield of floral leaves is increased, and the cultivation time is shortened; harvesting is easier after dwarfing, and the labor productivity is improved; when 3-4 pairs of true leaves grow on the cannabis sativa seedlings, cold treatment is carried out, at the moment, the seedlings mainly grow in a vegetative mode and do not grow in a reproductive mode, and the flower bud differentiation process is promoted to advance after cold treatment; a female inducing agent is sprayed to the cannabis sativa, so that the number of female flowers is greatly increased during sex differentiation of the cannabis sativa; a synthesis promoting agent is injected into the cannabis sativa, CBDA synthetase genes are increased, CBDA synthetase is increased, synthesis of CBD is promoted, and therefore the content of CBD is increased.

The invention provides a method for judging mauremys mutica embryonic development phase without dissection, and belongs to the field of temperature-dependent sex determination animal sex differentiation and sex regulation and control. In a mauremys mutica incubation season, mauremys mutica zygotes are collected for incubation; the change condition of fertilization spots is observed in the whole incubation process; the change condition of blood streaks is observed through light illumination and photos are taken for recording; in a first week of the incubation, the phase change is fast, and embryos are too small to be accurately judged by naked eye observation; embryo dissection observation is not performed in the first week of the incubation; 4 to 6 zygotes are taken every day for appearance observation from the eighth day of the incubation; then, the dissection is performed; the embryo development condition is observed in a stereoscopic microscope; text description and photo taking recording are performed; the embryonic development phase of Chinese softshell turtles is divided according to the division standard of the mauremys mutica embryonic development phase. The zygote dissection is not needed; through the appearance observation on the zygotes, the accurate development phase of the whole process before the starting of a mauremys mutica temperature sensitive period to the embryonic development completion can be obtained.

The invention provides a method for determining a sex differentiation phase of Scophthalmus maximus by means of high temperature induction. The method includes the steps of (1) building a full-sib family, (2) conducting a high temperature induction experiment and (3) determining the sex differentiation phase. According to the method, a control group and an experimental group where the other breeding conditions except water temperature and management are basically the same are set, larva fishes bred with the step (1) are stocked into water tanks of the experimental group and the control group respectively, the water temperature of the control group remains constant, a period of high temperature induction treatment is conducted on the experimental group at different developmental stages, and then cultivation is conducted on the experimental group and the control group where the water temperature remains the same until the sizes of fingerlings of the control group and the experimental group reach the sizes where sexes can be observed and identified through dissection; a relational graph of the male phenotype proportion of the experimental group and the fingerling size at the beginning of high temperature induction is drawn, wherein the fingerling size corresponding to the interval where a broken line of the male phenotype proportion changes obviously is the sex differentiation phase of Scophthalmus maximus. By means of the method, determination of the sex differentiation phase of Scophthalmus maximus is more accurate.

The invention discloses a siRNA-IR sequence of male Macrobrachium rosenbergii sex transition and its application; it uses siRNA interference technology to inhibit the expression of IR gene, promotes the sex transition of male Macrobrachium rosenbergii, and obtains pseudo-female shrimp; The IR gene fragment of Macrobrachium rosenbergii was screened and obtained from the transcriptome of Macrobrachium rosenbergii, and siRNA was designed and synthesized according to the IR gene sequence of Macrobrachium rosenbergii, named siRNA‑IR. Before the sex differentiation period, it was introduced into Macrobrachium rosenbergii by microinjection method, interfered and inhibited the expression of IR gene in Macrobrachium rosenbergii, and promoted the sex transition of Macrobrachium rosenbergii. Compared with dsRNA interference technology, siRNA interference technology has a smaller injection dose, can reduce costs, can effectively reduce the immune stress response of postlarvae of Macrobrachium rosenbergii, and improve the survival rate of pseudo female shrimp.