42 results about "Induced pluripotent stem cell" patented technology

Disclosed in the present invention are a biomimetic vascularized iPSC-hep spheroid for liver regeneration, and a preparation method therefor. The biomimetic vascularized iPSC-hep spheroid is a microcapsule with a porous hyaluronic acid methacryloyl as a shell and a human-induced pluripotent stem-cell-derived hepatocyte spheroid as an inner phase, and human umbilical vein endothelial cells adhere to the surface of the shell. The biomimetic vascularized iPSC-hep spheroid of the present invention promotes effective communication between hiPSC-heps and HUVECs, and provides barrier protection for an encapsulated hiPSC-hep spheroid so as to avoid attack by the immune system. Compared with dispersed cells, the cell-to-cell interaction in the cell spheroid improves cell-to-cell communication, helps to simulate the physiological heterogeneous structure and cell microenvironment of the liver, and can more effectively repair an acutely failing liver.

Provided are a method for inducing differentiation of pluripotent stem cells into CD34+ hematopoietic stem / progenitor cells and a culture medium composition thereof. Provided are a method for inducing differentiation of pluripotent stem cells into CD34+ hematopoietic stem / progenitor cells or NK cells and a culture medium composition thereof. The NK cells obtained using the provided culture method have high purity and good in vitro expansion effect; the yield of iNK cells is high, and a single iPSC can differentiate into approximately 2,000 NK cells; and the obtained iNK cells highly express CD16 (over 70%). As a result, the problem of low CD16 expression in iNK cells in the prior art, which requires genetic modification methods to solve, has been addressed.

The present invention relates to a method for producing adipocyte-derived dedifferentiated stem cells by using centrifugation. The method for producing adipose cell-derived dedifferentiated stem cells by using centrifugation, and adipose cell-derived dedifferentiated stem cells using same, according to one embodiment of the present invention, allow a production period to be significantly shorter than that of adipose cell-derived dedifferentiated stem cells produced using a conventional ceiling culture method, and thus can be effectively used in the field of autologous adipose-derived stem cell therapeutic agents.

[Summary] [Problems] The main purpose of this invention is to provide a technique for inducing an epidermal keratinocyte that can produce a three-dimensional cultured skin model with a multi-layered horny cell layer, from a pluripotent stem cell. [Solution] The present invention is a method for inducing differentiation from a pluripotent stem cell to an epidermal keratinocyte, using a non-detached cell after being treated with a cell detaching agent in the inducing step from the pluripotent stem cell, wherein when applying the epidermal keratinocyte to a three-dimensional cultured skin model, a three-dimensional cultured skin model in which the horny cell layer is multi-layered is obtained.

The present invention relates to a pharmaceutical composition for the prevention or treatment of epilepsy, and an epilepsy therapeutic agent screening method and a composition therefor, and specifically provides an epilepsy therapeutic agent screening method and an epilepsy therapeutic agent screening composition using induced pluripotent stem cells (iPSCs) derived from an epilepsy patient having a genotype comprising an SCN2A gene mutation, and accordingly, a pharmaceutical composition for the prevention or treatment of epilepsy having more excellent efficacy and selectivity than conventional epilepsy therapeutic agents may be screened and provided.

The invention provides a preparation method of adult T cells and universal adult CAR-T cells induced and differentiated by pluripotent stem cells, and belongs to the technical field of cell differentiation. By intervening the SP140 gene or SP140 protein, advanced hematopoiesis of pluripotent stem cells can be induced, and hematopoietic precursor cells and adult T cells with T gonadotrophy differentiation potential are obtained. The inducible CAR is knocked into the TRAC site of the pluripotent stem cell at a fixed point, so that endogenous regulation of CAR expression is realized, the in-vivo maintenance and anti-tumor capability of CAR-T is improved, and TCR-mediated graft versus host disease is blocked. The invention also provides a separated CAR-T cell, the CAR-T cell aiming at different antigens can be prepared by editing a seed cell of a pluripotent stem cell once, B2M and CIITA genes are further knocked out, HLA-E is overexpressed, and the immunogenicity of the CAR-T cell and the resistance of a host to the CAR-T cell are reduced.

The present invention relates to NK cells with an improved ability to target and kill cancer cells and use thereof. Specifically, the present invention relates to: iPSC-based NK cells into which a chimeric antigen receptor targeting a specific cancer antigen, CXCR4 and CCR7 genes, which are chemokine receptors, and an IL-15 / IL-15Rα (IL-15RF) gene are introduced; and use thereof for treating cancer. The iPSC-based NK cells according to the present invention can not only directly target a specific cancer antigen by the introduced CAR, but can also activate NK cells and enhance the proliferation thereof by regulating intracellular signaling and tumor microenvironment by the introduced CXCR4 and CCR7 and the introduced IL-15RF. Therefore, the NK cells into which multiple genes are introduced, according to the present invention, can efficiently target and attack cancer cells, and thus can be effectively used for the treatment of cancer diseases.

Described herein are methods and culture medium, useful for inducing polarization in multipotent stem cells. Additionally, described herein are multipotent cells produced by the methods and culture medium of this disclosure that are useful therapeutic agents. Also described herein are extracellular vesicles and factors secreted by multipotent cells that are produced by the methods and culture medium of this disclosure that are useful as therapeutic agents.

The invention discloses a kit and a method for constructing a human bone organoid from pluripotent stem cells, and relates to the technical field of organoid culture. The kit for constructing the human bone organoid by the pluripotent stem cells comprises a culture medium A, a culture medium B, a culture medium C, a culture medium D and a culture medium E. The culture medium A induces the pluripotent stem cells to form original stripes, the culture medium B induces the original stripes to form paraaxial mesoderm, the culture medium C induces the paraaxial mesoderm to form body wall mesoderm, the culture medium D induces body wall mesoderm cells to differentiate to bone cells, and the culture medium E induces bone cells to form cartilage organs. The kit provided by the invention can induce the pluripotent stem cells into bone organs, and has important significance in research and treatment of pathogenesis of bone related diseases and related research of bone repair and regeneration.

A composition for preventing or treating cancer or infectious disease is disclosed. The composition contains, as an active ingredient, a pluripotent stem cell-derived immune cell expressing IL-7, CCL19, or a combination thereof. The composition provides a multifaceted and synergistic therapeutic effect derived from the complementary immune response of the patient's endogenous T cells and the injected natural killer cells, by administering only a therapeutically effective amount of immune cells other than T cells, specifically natural killer cells. The natural killer cells of the composition may also be co-administered with exogenous T cells to allow these different cell populations to act more intensively at the lesion site. The composition is based on the differentiation of pluripotent stem cells, specifically induced pluripotent stem cells (iPSCs) into immune cells, thus can be used to generate an unlimited supply of allogenic or autologous cells as needed.

The present disclosure relates to methods and compositions for reprogramming cells to a pluripotent state. In particular, it relates to an integration- and feeder cell-free method for reprogramming primary human fibroblast cells to induced pluripotent stem cells (iPSCs).

The invention belongs to the technical field of stem cell induction, and discloses an epigenetic regulation-based iPSC (induced pluripotent stem cell) efficient directional induction method, which is characterized in that 5-azacytidine, vitamin C and alpha-ketoglutaric acid are added into somatic cells before induction so as to regulate DNA (deoxyribonucleic acid) methylation level and histone modification state, relieve an epigenetic inhibition structure and activate a pluripotent gene promoter region. Then, under the condition of no virus and no integrated vector, two specific micromolecules RG108 and MGCD0103 are introduced to respectively target DNA methyltransferase and histone deacetylase, and are matched with vitamin C to enhance TET enzyme activity, so that the somatic cells are efficiently induced to be reprogrammed into the induced pluripotent stem cells. The obtained iPSC has high clone formation rate, typical pluripotent marker expression and good genetic stability, has the advantages of high safety, high efficiency and short period, and is suitable for clinical research and individualized regenerative medicine application.

The present invention provides: a method for differentiating pluripotent stem cells into mesenchymal stem cells in the presence of a BMP4 signaling agonist; mesenchymal stem cells obtained thereby; and extracellular vesicles obtained from a culture solution of the stem cells. The extracellular vesicles have excellent anti-inflammatory effects, and thus can be used for treating inflammatory diseases. Therefore, there is a high possibility that a composition comprising the extracellular vesicles can be variously used as a pharmaceutical composition for treatment of inflammatory diseases and a cosmetic composition for skin improvement.

The present application provides a method for producing, from pluripotent stem cells, neural progenitor cells of the midbrain floor plate region or dopaminergic neural progenitor cells, the method comprising (1) inducing differentiation of pluripotent stem cells into neural progenitor cells of the midbrain floor plate region, and optionally (2) inducing differentiation of the neural progenitor cells of the midbrain floor plate region, which are obtained by the differentiation induced in (1), into dopaminergic neural progenitor cells, wherein all or part of the differentiation induced in (1) includes culturing the cells in a medium containing an agonist for LPA receptor.

In particular, compositions comprising iPSC-derived parietal cell precursors (iMPCs) generated using NK3 homeobox 1 (NKX3.1; parietal cell fate-determining transcription factor) and methods for their preparation and use are described herein. In addition, in particular, methods for maturing iMPCs into functional parietal cell subtypes, including smooth muscle cells, perivascular cells, and fibroblasts, as well as methods for increasing angiogenesis, angiogenesis, and cell junctions, are described herein. iMPCs mediate the formation of functional blood vessels when transplanted together with endothelial cells (ECs); thus, methods for modeling vascular diseases (e.g., 3D vascular organoids (VOs)) and therapeutic vascularization, comprising a method of administering iMPCs and ECs and a step of administering iMPCs and ECs, are also described herein.

This invention provides a gene therapy for Deutsche Syndrome (DSS) and relates to the field of clinical medical technology. It provides highly efficient pegRNA / ngRNA sequences and their application methods, validating the effectiveness of the lead editing system in treating SDS in two in vitro models: induced pluripotent stem cells (iPSCs) and hematopoietic stem / progenitor cells (HSPCs). Furthermore, the therapeutic effect of lead editing on SDS was identified in vivo by transplanting edited HSPCs, laying the foundation for clinical translation.

The present invention provides a method for producing an artificial bone-cartilage complex, the method comprising: (i) a step for inducing differentiation of pluripotent stem cells to obtain a precartilage cell mass; (ii) a step for seeding and culturing the precartilage cell mass obtained in the step (i) on the surface of an artificial bone having communication holes; and (iii) a step for recovering an artificial bone-cartilage complex in which a cartilage layer is formed on the surface of the artificial bone. Also provided is an artificial bone-cartilage complex produced by said method.

Provided is a technology allowing for stable supply of brain microvascular endothelium-like cells. This coating agent for inducing differentiation of pluripotent stem cells into brain microvascular endothelium-like cells contains at least one component of a Laminin-221 fragment or an N-terminal Vitronectin.

The invention discloses a method for generating human neural stem cells through IPS cell induction, and relates to the technical field of cell differentiation culture. The method comprises the following steps: (1) adding IPS cells into a culture medium I for culturing, and adding the IPS cells into a culture medium II for culturing to obtain cells A; (2) culturing the cell A in a culture medium III to obtain a cell B; (3) culturing the cell B in a culture medium IV to obtain a cell C; (4) culturing the cells C in a culture medium V to obtain neural stem cells; the culture medium III is prepared from a basic culture medium, 0.1 to 0.5 mM of L-glutamine, a transforming growth factor-beta with a concentration of 50 to 100 ng / ml, rhizoma panacis majoris saponin with a concentration of 1.5 to 5 ng / ml, magnolol with a concentration of 0.5 to 1 ng / ml and fetal calf serum with a concentration of 1 to 3 vol%. The method is simple, and directional differentiation of the IPS pluripotent stem cells to the neural stem cells can be induced under the synergistic effect of all the components in the culture medium.

To provide a novel method for producing iPS cells that requires less patient-derived somatic cells as raw material and reduces the use of reagents (e.g., viral vectors) compared to conventional methods. [Solution] The present invention provides a method for producing iPS cells using a cell processing device. The cell processing device has a closed system portion in which a sealed container for supplying materials and a centrifugal chamber configured to perform counterflow centrifugal separation are connected via a connecting conduit. The production method involves performing a step (s1) of contacting somatic cells with reprogramming factors while performing counterflow centrifugal separation in the centrifugal chamber, while maintaining the closed nature of the closed system portion.

Methods and compositions are provided for obtaining functionally enhanced derived effector cells obtained from directed differentiation of genomically engineered iPSCs. The iPSC-derived cells provided herein have stable, functional genome editing that provides improved or enhanced therapeutic effects. Therapeutic compositions and uses thereof are also provided, comprising functionally enhanced derived effector cells alone or in combination with antibodies or checkpoint inhibitors.

The invention belongs to the technical field of biology, and particularly relates to a method for inducing pluripotent stem cells to be differentiated into CD34 positive hematopoietic endothelial cells. According to the technical scheme, the method for inducing the pluripotent stem cells to be differentiated into the CD34 positive hematopoietic endothelial cells comprises the following steps: a, culturing the pluripotent stem cells until the convergence degree of the pluripotent stem cells reaches 50-80%; b, digesting the pluripotent stem cells into single cells, and resuspending the pluripotent stem cells into a culture medium to obtain a cell suspension; c, putting the cell suspension into a cell culture plate, and performing adherent culture; d, after the culture in the step c is completed, discarding the culture medium in the step b, adding a hematopoietic differentiation culture medium into the cell culture plate, and performing induced culture to form CD34 positive hematopoietic endothelial cells; the hematopoietic differentiation culture medium is composed of a D / F12 culture medium and salidroside. The invention is mainly used for providing a new choice for preparing hematopoietic endothelial cells.

Provided are methods for administering allogeneic induced pluripotent stem cells (iPSC)-derived midbrain dopaminergic progenitors to a subject for treatment of Parkinson's disease (PD). The midbrain dopaminergic progenitor cells are administered to 5 trajectories, each with 8 depositional sites, per putamen. Patients are operated on using a trans-frontal approach with an entry point near the coronal suture. Also provided are compositions and articles of manufacture for use in the methods.

The present invention provides a microfluidic system and method for reprogramming, proliferating, preserving, and optionally differentiating cells. [Solution] An integrated microfluidic system is provided that utilizes microfluidic chip technology to accept a patient sample containing cells, grow the cells, reprogram the grown cells, and then store the reprogrammed cells in a microfluidic chip. These microfluidic chips containing the reprogrammed cells can then be used in situations of genetic differentiation to specific cell types. Overall, this system and workflow is suitable as a hospital-based device that enables the creation of any patient-derived iPSC for downstream diagnostic or therapeutic applications.

Described herein are methods and culture medium, useful for inducing polarization in multipotent stem cells. Additionally, described herein are multipotent cells produced by the methods and culture medium of this disclosure that are useful therapeutic agents. Also described herein are extracellular vesicles and factors secreted by multipotent cells that are produced by the methods and culture medium of this disclosure that are useful as therapeutic agents.

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.

The invention discloses application of pluripotent stem cells to induced treatment of osteoarthritis, and belongs to the technical field of regenerative medicine. According to the research, a natural inducer is used for inducing pluripotent stem cells to be differentiated into mesenchymal stem cells, cartilage precursor cells and mature cartilage cells, and the mature cartilage cells and the mesenchymal stem cells are compounded to prepare the medicine for treating osteoarthritis to treat white rabbits with osteoarthritis, so that osteoarthritis inflammation can be remarkably eliminated, and damaged cartilage can be repaired; the curative effect is obviously superior to that of treatment by singly using mature cartilage cells or pluripotent stem cells. Through the synergistic effect of the mature cartilage cells, the human induced pluripotent stem cells, the natural inducer with the effects of diminishing inflammation and promoting cartilage regeneration, the dexamethasone and the glucosamine in combination with the collagen carrier, a microenvironment is provided for cell growth, the cell survival rate is guaranteed, inflammation is specifically inhibited, articular cartilage is repaired, and the cartilage repairing effect is improved. The invention provides a new scheme with better effect and more comprehensive mechanism for the treatment of osteoarthritis, and has important clinical transformation value.

The present invention provides methods for producing megakaryocytes and platelets derived from inducible pluripotent stem cells. Such megakaryocytes or platelets may be genetically modified to include a nucleic acid molecule encoding a therapeutic agent. The invention also provides methods and compositions for loading therapeutic agents to platelets or megakaryocytes and for genetically modifying platelets or megakaryocytes to express therapeutic agents.