19 results about "CD34" patented technology

The integration procedure provides different elements that require in vitro amplification of HSPCs, preferably for about 48 hours, in the presence of hematopoietic cytokines as well as UM171 and stem cell regenerative factor 1, and the introduction of i) a system (e.g., a CRISPR / Cas9 system) that generates DNA double-strand breaks (DSBs) upstream of the transcription start site of the target gene, and ii) the introduction of a donor matrix comprising cDNA with homologous arms on both sides (left [LHA] and right [RHA]).

本发明公开了一种CD34+造血干细胞化学重编程为诱导多能干细胞的方法。所述方法包括：将CD34+造血干细胞分散为单细胞，接种于包被细胞外基质的培养表面，在无血清培养基中依次进行三阶段诱导培养：第一阶段培养基包含丙戊酸、CHIR99021、RepSox、EPZ6438、PY60、OAC1和OAC2；第二阶段培养基包含丙戊酸、CHIR99021、RepSox、反苯环丙胺、TTNPB、Forskolin、OAC1和OAC2；第三阶段培养基包含CHIR99021、Forskolin、Y‑27632、SP600125、OAC1和OAC2。最后挑取形成的iPSC克隆进行扩增培养，获得诱导多能干细胞。本发明针对CD34+造血干细胞优化了小分子组合和诱导时序，全程无病毒、无转录因子、无血清，重编程效率可达3%以上，细胞凋亡率低，可获得具有完整多能性和正常核型的iPSC，适用于临床级细胞制备和再生医学应用。

This invention relates to an immunoRaman multicolor imaging kit for the prediction and assessment of liver cancer. The immunoRaman multicolor imaging reagent contains five Raman probes, which are composed of Raman spheres and antibodies attached to the surface of the Raman spheres. The Raman spheres are polymerized from triple-bonded monomers with (i) acrylic acid or (ii) styrene and acrylic acid, with a particle size of 10–100 nm and a Raman shift of 1800–2800 cm⁻¹. ‑1 The antibodies used for the five Raman probes are selected from one of the following: AFP antibody, HBX antibody, GPC3 antibody, GS antibody, and CD34 antibody. The immunoRaman multicolor imaging reagent and kit provided by this invention can achieve one-step labeling and multiplex color development of five liver cancer-related protein markers in the same liver tissue section. Compared with multiplex fluorescence immunohistochemistry, this invention has significant advantages in both efficiency and accuracy. Experiments show that the detection results of this invention are consistent with clinical diagnostic results, demonstrating great application potential.

Described herein are compositions and methods for modifying eukaryotic cells, for example, to express a transgene of interest and / or to produce an expanded population of cells ex vivo. Using the compositions and methods of the disclosure, a population of eukaryotic cells, such as a population of pluripotent cells (e.g., CD34+ hematopoietic stem or progenitor cells) may be transduced to express a gene of interest by contacting the cells with a viral vector, such as a lentiviral vector, and a poloxamer. Additionally, the compositions and methods described herein can be used to promote the proliferation or survival of a population of pluripotent cells (e.g., CD34+ hematopoietic stem or progenitor cells) ex vivo, for example, by contacting the cells with a poloxamer. Examples of poloxamers that may be used in conjunction with the compositions and methods of the disclosure are those having a molar mass in excess of 10,000 g / mol, as well as those having a molar mass of polyoxypropylene subunits greater than 2,000 g / mol and / or an ethylene oxide content of greater than 40% by mass.

Methods to improve homing and engraftment of hematopoietic stem cells, particularly cord blood CD34+ cells, for use in hematopoietic cell transplantation through the regulation of expression of YTHDF2 or FTO in CD34+ cells. The methods include transiently repressing expression of YTHDF2 through exposure of the CD34+ cells to YTHDF2 repressor compound. The methods also include exposing CD34+ cells to a FTO expression activator compound to transiently increase FTO expression in the cells.

Mastocytosis is a rare and heterogeneous disorder marked by the abnormal accumulation of mast cells (MCs) in various tissues. Tyrosine kinase inhibitors have shown limited efficacy, suggesting that additional molecular mechanisms are involved. The inventors showed the unexpected expression of AXL in neoplastic mast cells from patients with various forms of mastocytosis. This is particularly noteworthy as AXL expression had never been investigated in mast cells, whether neoplastic or normal. The study demonstrated the role of AXL, particularly in cooperation with KIT D816V to drive mast cell proliferation and resistance to therapies. In an embodiment, the AXL inhibitor bemcentinib (R428) is used in combination with the TKI midostaurin (PKC 412). The AXL-L197M identified in the ASM patient is pro-proliferative, and induces resistance to death in CD34-derived mast cells and in Ba-f3. Furthermore, the results also suggest that a combination of AXL inhibitors and BCL-2 inhibitors may be appropriate for the treatment of mastocytosis. In conclusion, AXL inhibitors can be thus suitable for the treatment of mastocytosis.

This invention provides the application of T-5224 in the expansion and / or functional enhancement of human circulating hematopoietic stem / progenitor cells, relating to the fields of stem cell biology and regenerative medicine. It is the first time T-5224 has been applied to the expansion of cHSPCs, opening up a new pathway to improve stem cell culture efficacy by regulating downstream transcriptional hubs of inflammatory / stress signals. Accompanied by the upregulation of HOXB4 gene expression, it significantly improves LIN... ‑ CD45RA ‑ CD34 + CD38 low / ‑ The cell expansion factor, maintaining cell stemness and regeneration capacity, provides a new approach to solving the technical problem of the difficulty in rapidly expanding human circulating hematopoietic stem / progenitor cells in in vitro culture while maintaining cell stemness and regeneration potential.

A non-invasive method for detecting bone marrow pathology is provided, comprising receiving a metacell model of multiple metacell types based on single-cell RNA sequencing (scRNA-seq) of CD34-positive cells from peripheral blood and comparing it to a control value of CD34-positive cell metacells from peripheral blood of a healthy subject. A non-invasive method for predicting the percentage of blast cells in the bone marrow and calculating an IPSS-M risk score is also provided, as well as a system for carrying out the method of the present invention.

The application discloses an optimized method for preparing amniotic membrane stem cell membrane pieces, relates to the technical field of tissue engineering and regenerative medicine, and significantly shortens the preparation period of the amniotic membrane stem cell membrane pieces by adding nicotinamide adenine dinucleotide (NAD+) and a precursor compound beta-nicotinamide mononucleotide (NMN) in a traditional cell membrane piece culture medium, combining with vitamin C (VC) and a mechanical scratching method. More importantly, the method can effectively reduce the surface expression amount of stem cell CD34 in the membrane pieces, from about 2.04% in a conventional method to below 0.12%, and the prepared stem cell membrane pieces have lower immunogenicity and more definite mesenchymal stem cell characteristics, so that the safety and effectiveness of the stem cell membrane pieces in clinical transplantation application are greatly improved.

This invention provides the application of phloroglucinol in the expansion and / or functional enhancement of human circulating hematopoietic stem / progenitor cells, relating to the fields of stem cell biology and regenerative medicine. Phloroglucinol possesses dual inhibitory capabilities against both NF-κB and JNK / c-Jun, effectively reducing the protein level of phosphorylated NF-κB p65 (Ser536) in cultured cells, accompanied by upregulation of HOXB4 gene expression, and significantly improving LIN... ‑ CD45RA ‑ CD34 + CD38 low / ‑ The expansion fold of cHSPCs cells, maintaining cell stemness and regeneration capacity, provides a new approach to solving the technical problem of the difficulty in rapidly expanding human circulating hematopoietic stem / progenitor cells in in vitro culture while maintaining cell stemness and regeneration potential.

This invention discloses a detection kit and method for abnormal lymphocytes, myeloid blast cells, and acute promyelocytic leukemia. The detection kit includes a first reagent and a second reagent. The first reagent includes a hemolysin and a staining agent. The second reagent includes a hemolysin, a staining agent, and an antibody treatment agent, including CD13 and CD34 antibody treatment agents. The hemolysin, based on a total volume of 1L, includes the following components: 0.1-10g castor oil polyoxyethylene ether nonionic surfactant, 0.01-10g cationic surfactant, 0.5-15g buffer, 0.01-8g preservative, with the remainder being water. The pH value of the hemolysin is 5-9. The staining agent, based on a total volume of 1L, includes the following components: 0.001-0.2g nucleic acid dye, with the remainder being a solvent. The detection method includes: obtaining a blood sample to be tested, preparing a first sample, testing the first sample, preparing a second sample, and testing the second sample. This invention can accurately detect abnormal lymphocytes, myeloid blast cells, and diagnose acute promyelocytic leukemia.

This invention relates to a method for promoting somatic cell reprogramming to generate iPSC clones. Specifically, it provides a small molecule composition for rapidly promoting CD34+ blood cell reprogramming to generate iPSC clones.

This invention relates to the field of hematopoietic stem cell technology, specifically to a culture medium combination and method for inducing pluripotent stem cells to differentiate into hematopoietic stem cells. Addressing the problems of low efficiency and low yield associated with existing methods that require digestion and sorting for pluripotent stem cell differentiation into hematopoietic stem cells, resulting in only one harvest after induction, this invention provides a culture medium combination and method that enables "one-time induction, multiple harvests." This invention adds CHIR99021 to culture medium I and SB431542 to culture medium III, combined with other culture medium components and dynamic culture, to achieve highly efficient mesoderm induction. Combined with periodic replacement of culture medium V, digestion and sorting are unnecessary, allowing for more than three periodic harvests of high-purity (CD34) stem cells in the same container from a single induction. + CD45 + >95% hematopoietic stem cells. This invention simplifies the operation steps, eliminating the need for re-seeding, digestion, or sorting and enrichment, significantly increasing the yield per batch and making it suitable for industrial production; the entire process is free of foreign substances, meets GMP standards, and the resulting cells meet clinical application requirements.

This invention provides the application of JSH-23 in the expansion and / or functional enhancement of human circulating hematopoietic stem / progenitor cells, relating to the fields of stem cell biology and regenerative medicine. The small molecule compound JSH-23 effectively reduces the protein level of phosphorylated NF-κB p65 (Ser536) in cultured cells, accompanied by upregulation of HOXB4 gene expression, and significantly improves LIN. ‑ CD45RA ‑ CD34 + CD38 low / ‑ The cell expansion factor, maintaining cell stemness and regeneration capacity, provides a new approach to solving the technical problem of the difficulty in rapidly expanding human circulating hematopoietic stem / progenitor cells in in vitro culture while maintaining cell stemness and regeneration potential.

Embodiments of this disclosure relate to methods and compositions relating to the optimization and selection of umbilical cord blood units for the production of immune cells, such as natural killer (NK) cells, for use in adoptive cell therapy. In certain embodiments, specific properties of an umbilical cord blood unit and / or the properties of cells obtained therefrom are analyzed. If a threshold measurement is met for one or more properties of the umbilical cord blood unit and / or the properties of cells obtained therefrom, the umbilical cord blood unit is used as a source for the production of immune cells. Specific properties to be measured include, for example, the viability of umbilical cord blood cells, the time from birth to cryopreservation of the umbilical cord blood unit, the total nucleated cell recovery rate, infant weight, infant sex, maternal age, gestational age, CD34-positive cell percentage, and / or nucleated red blood cell content. These properties may be determined before and / or after cryopreservation.

The application discloses a chimeric antigen receptor targeting CD180 and application thereof. Through flow cytometry, degranulation analysis experiment and detection of T cell secreted cytokines, it is proved that the T cell modified by the chimeric antigen receptor has a strong killing effect on acute myeloid leukemia cells expressing CD180, has no killing effect on cells not expressing CD180, effectively prevents off-target effect, and does not affect the colony formation ability of umbilical cord blood derived CD34 + hematopoietic stem / progenitor cells, and is safe. The chimeric antigen receptor CD180scFv-CD8alpha-4-1BB-CD3zeta of the application can be used for treating CD180 positive blood tumors.

A novel method for obtaining NK cells, which includes culturing CD34+ cells in the presence of TNF-α or of a fragment thereof and of a Notch ligand or fragment thereof, thereby obtaining a first population of cells, and culturing the population of cells obtained in a cytokine-containing medium. Also, a NK cell population, and the use thereof for treating diseases, including cancer.

The application relates to the technical field of cell culture, and discloses an in-vitro expansion method of CD34+ hematopoietic stem cells, which comprises the following steps: inoculating separated CD34+ cells into an expansion culture medium which is completely free of exogenous recombinant cytokines and animal-derived albumin for culture; wherein the expansion culture medium comprises: a) a basic culture medium; b) a chemical agonist combination comprising a phosphoinositide 3-kinase activator, a thrombopoietin receptor agonist and a pyrimidine indole derivative UM171; and c) a synthetic polymer. The application provides an in-vitro expansion method of CD34+ hematopoietic stem cells, and provides a novel culture method which can get rid of dependence on exogenous complex biological products, has clear chemical components and can specifically support functional HSCs self-renewal and non-differentiation expansion.