103 results about "Trophoblastic cell" patented technology

The present invention provides methods and compositions for specifically identifying a fetal cell. An initial screening of approximately 400 candidate genes by digital PCR in different fetal and adult tissues identified a subset of 24 gene markers specific for fetal nucleated RBC and trophoblasts. The specific expression of those genes was further evaluated and verified in more defined tissues and isolated cells through quantitative RT-PCR using custom Taqman probes specific for each gene. A subset of fetal cell specific markers (FCM) was tested and validated by RNA fluorescent in situ hybridization (FISH) in blood samples from non-pregnant women, and pre-termination and post-termination pregnant women. Applications of these gene markers include, but are not limited to, distinguishing a fetal cell from a maternal cell for fetal cell identification and genetic diagnosis, identifying circulating fetal cell types in maternal blood, purifying or enriching one or more fetal cells, and enumerating one or more fetal cells during fetal cell enrichment.

The invention provides for the production of several humanized murine antibodies specific for the antigen LK26, which is recognized by the murine antibody LK26. This antigen is expressed in all choriocarcinoma, teratocarcinoma and renal cancer cell lines whereas it is not expressed on cell lines of leukaemias, lymphomas, neuroectodermally-derived and epithelial tumour cell lines (excepting a small subset of epithelial cell lines). Furthermore, whereas renal cancer cell lines express the LK26 antigen, normal renal epithelial cells do not. Similarly, with the exception of the trophoblast, all normal adult and fetal tissues tested are negative for the LK26 phenotype. The invention also provides for numerous polynucleotide encoding humanized LK26 specific antibodies, expression vectors for producing humanized LK26 specific antibodies, and host cells for the recombinant production of the humanized antibodies. The invention also provides methods for detecting cancerous cells (in vitro and in vivo) using humanized LK26 specific antibodies. Additionally, the invention provides methods of treating cancer using LK26 specific antibodies.

Existence of human trophoblast stem (hTS) cells has been suspected but unproved. The isolation of hTS cells is reported in the early stage of chorionic villi by expressions of FGF4, fgfr-2, Oct4, Thy-1, and stage-specific embryonic antigens distributed in different compartments of the cell. hTS cells are able to derive into specific cell phenotypes of the three primitive embryonic layers, produce chimeric reactions in mice, and retain a normal karyotype and telomere length. In hTS cells, Oct4 and fgfr-2 expressions can be knockdown by bFGF. These facts suggest that differentiation of the hTS cells play an important role in implantation and placentation. hTS cells could be apply to human cell differentiation and for gene and cell-based therapies.

A human corneal limbal stem cell-amnion tissue engineering complex using human corneal limbal fibroblasts as the trophoblast is characterized in that: it is close to the human corneal epithelium form in histology form, and is analogous to the human corneal epithelium in immunology phenotype, its ultrastructure shows that the connection between the cells grows mature and the connection between the epithelia and the amnion carrier is a close conglutination. The culture method includes the following steps: (1) preparing the amnion for culturing the human corneal limbal stem cell in vitro; (2) preparing the human corneal limbal fibroblast trophoblast; (3) preparing the human corneal epithelia suspension; (4) culturing the human corneal limbal stem cell-amnion complex. The human corneal limbal stem cell-amnion complex has analogous phenotype to the human corneal epithelium, and provides a safe and effective biology tissue engineering product to treat corneal limbal stem cell defective eye diseases.

The invention provides a method for culturing human induced pluripotent stem cells (iPSCs) by using human bone marrow mesenchymal stem cells as a trophoblast. The method comprises the following steps of: obtaining the human iPSCs from child foreskin fibroblasts by using third to fifth generation human bone marrow mesenchymal stem cells obtained through subculture and culture after thawing from low-temperature freezing, performing amplification culture, and inoculating into trophoblast cells to obtain the human iPSCs. In the method, the human iPSCs are cultured by using the human bone marrow mesenchymal stem cells as the trophoblast cells instead of mouse embryonic fibroblasts in the conventional method, so that the pollution of heterologous cells in a culture system is reduced, that the culture system can make the human iPSCs amplified in vitro is proved, biological characteristics and multipotency of the human iPSCs are maintained for a long time, and the possibility of clinical application of the human iPSCs is provided.

The invention discloses a human umbilical cord blood hematopoietic stem cell (HSC) high-efficiency in vitro amplification technology. According to the invention, umbilical cord mesenchymal stem cells (MSCs) and umbilical cord blood plasma are used in combination for carrying out HSC in vitro amplification. A process comprises steps that: umbilical cord MSCs are cultured as a trophoblast; a special culture solution containing umbilical cord blood plasma is adopted as an amplification medium of the HSCs; and umbilical cord blood karyotes are adopted as amplification initiator cells. According to the invention, exogenous cell factors are not required to be added, treatments such as mitomycin C or gamma-ray 21Gy irradiation are not required by the trophoblast, no 3-dimensional technology is required, and the HSC high-efficiency amplification can be carried out. The technology is advantaged in low cost, convenient amplification, and low product immunogenicity. The technology is suitable for large-scaled productions. The practical application of the technology plays an important role in clinical treatments and researches.

The invention relates to a preparation method used for high efficiency amplification of natural killer (NK) cells. The preparation method comprises following steps: peripheral blood is collected, andsingle karyocytes are separated; the karyocytes are introduced into a culture bottle treated via coating with CD16 monoclonal antibody and HER2 monoclonal antibody for culturing; IL-2, IL-15, IL-21, and inactivated autologous serum are introduced into a culture medium at the same time; the cells are transformed into a culture bottle free of coating treatment, liquid supply is carried out every 2 to 3 days; and at last the obtained NK cells are collected. According to the preparation method, no heterogenous serum or trophoblastic cell is adopted, operation process is simple, high NK cell yieldis achieved, and the clinical application value is high.

A process for separating and culturing human epidermal stem cells includes such steps as providing ectocytic matrix covering Petri dish, separating epidermal cells, preparing trophoblastic cells and screening and culturing epidermal stem cells.

The invention relates to an integrated detection reaction orifice plate and protein chip reagent box for detecting six sterility indexes. And the reaction orifice plate comprises a substrate and reaction orifices on the substrate and the reaction orifices comprise 3-384 sample orifices, 1-300 negative contrast orifices, and 1-300 positive quality control orifices, where at the bottom of each reaction hole is solid carrier, coated with micro lattice of Sperm antigen (SAg), Zona pellucida antigen (ZPAg), Endometrium antigen (EMAg), Ovary antigen (OAg), Trophoblast antigen (TAg), and Human Chorionic Gonadotropin antigen (HCGAg) antigens or more. And the reaction orifice plate and reagent can simply and conveniently, rapidly and accurately implement simultaneously detection of six sterility indexes for many persons.

The invention discloses a conditional Cas9 expression induced swine trophoblastic cell line and an establishment method and application thereof. The method includes: subjecting a lentiviral vector, for controlling Cas9 gene expression, of a Tet-on system to virus packaging, concentrating and purifying; incubating lentivirus with a typical swine trophoblastic cell line, changing fluid, adding puromycin for drug screening, diluting cells to single ones, and culturing to obtain the single-cell-source swine trophoblastic cell line with Cas9 expression under Tet-on regulation. The swine trophoblastic cell line is simple in operation and convenient to use and has a potential utilization value in researching of key functional genes of some trophoblast cells and screening of swine gene target spots. The cell line is expected to be a significant cell material for researching swine placenta development and applicable to correlation researches on swine early-stage placenta development and uterine pregnancy mechanisms and also has a high reference value in research of human early embryonic development.

Methods for isolating and purifying fetal trophoblasts from a mucus sample obtained from the uterine cavity of a pregnant female. The mucus sample is transported from a clinical collection facility to a laboratory in a transportation medium so the cells remain viable. The mucus sample is then subjected to precise processing steps, including treatment with mucolytic agents or mucinases, sugar hydrolysis enzymes, nucleases, and proteases to provide fetal cells, the outer surfaces of which are so essentially completely devoid of attached mucosal biological material that they are then isolated in greater numbers than previously had been possible. The isolated cells are in appropriate condition to immediately be effectively subjected to FISH or to other molecular diagnostics.

The invention discloses a differential staining method for inner cell mass cells and trophoblastic cells of cattle blastulae. The method comprises the following steps: 1, carrying out immune combination on an anti-CDX2 monoclonal antibody which is adopted as a primary antibody and CDX2 molecules which are specifically expressed in the trophoblastic cells of the cattle blastulae through an immunostaining process; 2, the cattle blastulae which are cleaned and are combined with the primary antibody are immunostained with an antibody which is labeled by red fluorescence and can combine with the anti-CDX2 monoclonal antibody as a secondary antibody through the immunostaining process; and 3, whole nuclei of the cattle blastulae are subjected to fluorescent staining after completing the immunostaining to form the differential staining. On the basis of a case that CDX2 proteins specifically express in the trophoblastic cells and do not express in the inner cell mass cells, the method of the invention allows the differential staining to be carried out based on the red immunostaining of the CDX2 molecules and the blue staining of DAPI nuclei, so the accuracy is 100%, and obtained pictures are intelligible and beautiful.

The invention provides serum-based, diagnostic, biological assays for predicting disorders of pregnancy resulting from poor trophoblast and/or placental ischemia, including preeclampsia. Serum samples from such subjects exhibit an ability to disrupt the architecture involving fetal trophoblasts and maternal endothelial cells in a three-dimensional, dual cell co-culture system provided herein, in contrast to normal pregnancy serum samples. Based on these distinctions, the assays are employed to predict pregnancy outcomes as early as first trimester.

The invention discloses the application of Activin A for cell culture in human blast stem cell panoistic layer, belonging to biomedical technology field. It is necessary and adequate for Activin A to maintain self- refresh and multi-function for human blast stem cell, it can induce expression of transcription factor Oct4 and Nanog. The stem cell still possess differentiation potence in mouse body for teratoma after 10- generation culture with panoistic cell culture medium containing 5 ng/ ml Activin A, and it still maintains normal karyotype after 150- day culture and more than 20- generation.

A purified preparation of primate embryonic stem cells is disclosed. This preparation is characterized by the following cell surface markers: SSEA-1 (−); SSEA-4 (+); TRA-1-60 (+); TRA-1-81 (+); and alkaline phosphatase (+). In a particularly advantageous embodiment, the cells of the preparation are human embryonic stem cells, have normal karyotypes, and continue to proliferate in an undifferentiated state after continuous culture for eleven months. The embryonic stem cell lines also retain the ability, throughout the culture, to form trophoblast and to differentiate into all tissues derived from all three embryonic germ layers (endoderm, mesoderm and ectoderm). A method for isolating a primate embryonic stem cell line is also disclosed.

The invention belongs to the technical field of primary cell culture, and especially relates to a B lymphocyte in vitro culture system and applications thereof. The culture system utilizes a T lymphocyte line capable of secreting interleukin molecules such as IL-2 as trophoblastic cells so as to provide micro interleukin molecules required by the growing development of B lymphocyte; and trimer CD40L molecules expressed in vitro through a recombinant manner and having high biological activity are added as important molecules so as to further stimulate the growth, reproduction and development ofthe B lymphocyte. The system can also perform gamma ray irradiation treatment on the trophoblastic cells, so that the pre-treated trophoblastic cells can lose the ability of cell reproduction but still has the ability of protein synthesis and secretion within 7-14 days. The provided system can effectively culture the B lymphocyte derived from rabbits and alpacas in vitro, and can utilize the method to develop corresponding monoclonal antibodies.

The invention discloses a method for efficiently separating mouse spermatogonial stem cells. The method includes the following steps that firstly, mouse testis are collected, and a testicular cell suspension is prepared through a two-step enzyme digestion method; secondly, the testicular cell suspension is inoculated in a porous plate according to a certain density, and a spermatogonial stem cell culture solution is added for in-vitro culture; thirdly, the culture solution is removed, testis cells of different times are cultured through tryptic digestion, and spermatogonial stem cells are sorted from the testis cells through the magnetic activated cell sorting; fourthly, the sorted spermatogonial stem cells are inoculated to an STO trophoblast, and a spermatogonial stem cell culture solution is added for primary culture and subculture of the spermatogonial stem cells. A mouse spermatogonial stem cell system is built through the method including the steps of separation, culture, re-separation and re-culture. The method can achieve the aim of efficiently enriching the spermatogonial stem cells in vitro, and is economical and easy to implement, so that a direction is provided for protection of rare animal varieties and treatment of male infertility.

The invention relates to human placenta trophoblastic cell series which is conserved in China Committee for Culture Collection of Microorganisms on September 22th in 2005 with CGMCC No.1461 conserving number. It includes the following steps: separating the trophoblastic cell form the placenta tissue which is pregnant for 6-7 weeks; filtering the cells with multiplication capacity; processing subunit gene transfection by telomere enzyme to gain four long life cell strains. The cell series has normal multiplication capacity and biological chemical function, active cell growth, and reaches the 210 generation. Thus it can be used as target cell model for filtering antifertility drug and curing pregnant correlative disease medicine, provide ideal vitro model for further studying trophoblastic cell, lay important foundation for studying embryo implantation and pregnant physiology and pathological mechanisms.