15 results about "Limbal stem cell" patented technology

The invention provides a nerve growth factor functionalized exosome as well as a preparation method and application thereof. The functionalized exosome can activate a Wnt / beta-catenin signal channel to maintain dryness of corneal limbal stem cells (LSCs) so as to promote corneal epithelium repair, can activate a cAMP signal channel to relieve inflammatory response and promote corneal nerve regeneration, and can be used for treating or relieving corneal chemical burns, including relieving inflammatory response, reducing stromal scar formation and inhibiting corneal neovascularization. Corneal epithelium repair and nerve regeneration are promoted.

The invention provides a corneal limbus stem cell in-situ repair microneedle array patch and a preparation method and application thereof.The corneal limbus stem cell in-situ repair microneedle array patch comprises a biocompatible backing layer and a microarray layer arranged on the biocompatible backing layer, microneedles of the microarray layer are of a conical structure, the height is 100-300 micrometers, the vertex angle is smaller than 30 degrees, and the thickness of the microneedles of the microarray layer is 10-30 micrometers. And a nano-scale porous structure for loading cornea stem cell suspension is arranged in the cornea stem cell support. Through precise geometric design, biomimetic material selection and spatial arrangement optimization, in-situ delivery, microenvironment activation and tissue fusion in a corneal limbus area are realized under the condition of not depending on in-vitro amplification and surgical suture, and the method becomes an important direction for promoting LSCD minimally invasive treatment strategy innovation.

The present application provides a method for in vitro purified corneal limbal stem cells to be transdifferentiated into lens spherules, specifically, the present application inoculates corneal limbal stem cells onto a culture plate containing a Matrigel glue layer, and cultures the corneal limbal stem cells in a culture system containing Y-27632 additives in LSCM culture solution, so that lens spherule organoids can be obtained. The method of the present application can realize the rapid transdifferentiation of corneal limbal stem cells into lens spherules, so that they can be implanted into the capsular bag as seed cells for in situ regeneration of the lens.

The application discloses application of a norepinephrine inhibitor in preparation of a medicine for treating or relieving corneal limbal stem cell deficiency, and belongs to the technical field of medicines.The experiment proves that sympathetic nerves are excessively activated in a mouse model of corneal limbal stem cell deficiency, and the release of norepinephrine is increased, and the experiment proves that 6-hydroxydopamine hydrochloride can destroy sympathetic nerves to inhibit the release of norepinephrine, and a tyrosine hydroxylase inhibitor can inhibit the synthesis of norepinephrine, and both can further relieve the symptoms of corneal limbal stem cell deficiency.The norepinephrine inhibitor can improve the corneal limbal microenvironment, reduce corneal neovascularization, reduce corneal epithelialization, enhance corneal transparency and relieve corneal edema, and provides a new direction and a new way for the pathogenic mechanism and intervention target of corneal limbal stem cell deficiency, and has a wide application prospect.

The invention provides application of Mafb as a target spot in promotion of diabetic corneal epithelium injury repair, and belongs to the technical field of biological medicine. According to the application, the Mafb gene is taken as a treatment target, the repair speed of corneal epithelium injury can be obviously accelerated by over-expressing the Mafb gene in the cornea of a diabetic patient, and the expression levels of stem cell markers and cell proliferation markers in corneal epithelium tissues are obviously up-regulated. The invention also provides a recombinant adeno-associated virus AAV9-Mafb for overexpressing the Mafb gene, and the AAV9-Mafb can significantly recover the function of the corneal limbal stem cells of the diabetic mouse and promote the proliferation of corneal epithelial cells, thereby accelerating the repair of corneal epithelium injury of the diabetic mouse.

The invention belongs to the technical field of biology, and particularly relates to a cytoskeleton for promoting proliferation of corneal limbal stem cells. The raw materials of the cytoskeleton provided by the invention are chitosan and sodium hyaluronate, and the cytoskeleton has good biocompatibility and biological activity, can simulate the characteristics of an extracellular matrix, provides necessary physical and chemical support for stem cells, and does not cause immune response or cytotoxicity. The addition of the magnetic nanoparticles further enhances the performance of the cytoskeleton. Directional arrangement under the action of a magnetic field can provide structural support closer to an in-vivo microenvironment for cells, and adhesion and proliferation of the cells are promoted. The cell scaffold provided by the invention can provide a good growth microenvironment for the corneal limbus stem cells, and is beneficial to solving the problem that the corneal limbus stem cells are easy to lose activity and function in in-vitro culture.

The invention relates to corneal adhesive lipid nanoparticles based on mRNA (messenger ribonucleic acid) as well as a preparation method and application thereof, and belongs to the technical field of genetic engineering of ophthalmic related diseases. The preparation method comprises the following steps: preparing a linearized ID3 or PAX6 DNA fragment; preparing a linearized single chain ID3 or PAX6 mRNA (messenger Ribonucleic Acid) solution; and the mRNA-based corneal adhesive lipid nanoparticles are prepared by a self-assembly method. The lipid nanoparticles are prepared by the above method. The lipid nanoparticles are applied to preparation of a medicine for treating corneal epithelium injury. The method has the advantages that ID3mRNA or PAX6mRNA is successfully delivered to the corneal limbus stem cells, the dryness and self-renewal capacity of the corneal limbus stem cells after alkali burn or other injuries are quickly and efficiently recovered, and the multiplication capacity of the corneal limbus stem cells is promoted; the residence time of mRNA lipid nanoparticles on the corneal ocular surface can be prolonged, and the toxicity and immunogenicity are extremely low.

A method for constructing a graded limbal injury and repair model comprises: after administering topical anesthetic drops to the operated eye of a model animal, placing a limbal injury device containing a NaOH solution in the limbal area of ​​the model animal for 30 seconds, then removing the device and irrigating the operated eye with saline; postoperatively, administering an antibacterial drug to the operated eye of the model animal several times daily; and eight days after surgery, a graded limbal injury and repair model is obtained, wherein the limbal injury device applies alkali burns of varying sizes to the limbal area, thereby achieving a graded injury effect. The construction method provided by the present invention has been shown to be simple, laboratory-friendly, inexpensive, short, and reproducible, with quantifiable evaluation methods, making it an ideal method for constructing an animal model of limbal stem cell injury.

The invention relates to the technical field of veterinary ophthalmology, in particular to a cat corneal limbal stem cell immortalized cell line FLSC-SV40LT as well as a construction method and application thereof. The immortalized cell line FLSC-SV40LT is preserved in the China General Microbiological Culture Collection Center (CGMCC); the preservation number of the strain is CGMCC No. 46589; carrying out isolated culture on primary cat corneal limbal stem cells FLSC; immortalization treatment: infecting the primary FLSC cell line by using a lentivirus plasmid vector containing SV40LT; after infection, replacing a puromycin-containing BM culture medium for screening; screening survival cells, and carrying out multiplication culture, so as to obtain the immortalized cell line FLSC-SV40LT. Primary FLSCs are successfully separated, cultured and identified from cat corneal limbus tissues, SV40LT is used for successful lentivirus infection of the FLSCs, and a cat corneal limbus stem cell immortalized cell line is obtained for the first time and is named as FLSC-SV40LT; the establishment of the immortalized cell line FLSC-SV40LT provides an important basis for the basic research and clinical application of FLSC, and also provides an important research basis for veterinary ophthalmology, especially for cat corneal diseases, and comparative ophthalmology.

The invention discloses an application of PDGF-C in corneal injury repair. Research finds that in a corneal injury model mouse with Pdgf-c knockout, the corneal healing area in the corneal injury mouse model is reduced through Pdgf-c knockout, and the number of corneal limbal stem cells with proliferative activity is remarkably smaller than that of wild type mice. Through AAV-mediated PDGF-C overexpression, proliferation of corneal limbal stem cells can be significantly promoted, and the corneal healing area in a corneal injury mouse model is significantly increased. The results show that overexpression of the PDGF-C significantly improves the healing ability to corneal injury. Therefore, the invention provides a novel method for treating corneal injury.

The invention relates to hydrogel for corneal limbal stem cell transplantation and a preparation method thereof. The formula of the hydrogel comprises a polyether polymer, a gelatin derivative and a photoinitiator. The mass percentage content of the polyether polymer in the hydrogel is 1-5%; the mass percentage content of the gelatin derivative is 2-15%; the mass percentage content of the photoinitiator is 0.01-0.05%; the preparation method of the hydrogel comprises the following steps: (1) respectively preparing a polyether polymer solution, a gelatin derivative solution and a photoinitiator solution; and (2) mixing the three solutions according to a certain volume ratio, and uniformly stirring to obtain the product. Wherein the volume ratio is (300-500): (200-1000): (1-5). Aiming at the problems of single carrier structure, poor transplantation efficiency, limited tissue repair performance and the like in the current clinical corneal limbal stem cell transplantation system, the invention firstly proposes that the modified hydrogel can be used for treating ocular surface diseases and a corneal limbal stem cell-loaded hydrogel transplantation system is constructed for the first time, so that the corneal limbal stem cell-loaded hydrogel transplantation system can be used for preparing the corneal limbal stem cell-loaded hydrogel transplantation system. An efficient transplantation strategy matched with the method is provided, and repairing and remodeling of the ocular surface are effectively promoted.

The application discloses application of PDGF-C in corneal injury repair. The application research finds that in a corneal injury model mouse in which Pdgf-c is knocked out, Pdgf-c knockout reduces the area of corneal healing in the corneal injury mouse model, and the number of corneal limbal stem cells with proliferative activity is significantly less than that of wild type mice. Through AAV-mediated PDGF-C overexpression, the proliferation of corneal limbal stem cells can be significantly promoted, and the area of corneal healing in the corneal injury mouse model is significantly increased. These results show that overexpression of PDGF-C significantly improves the ability of corneal injury healing. Therefore, the application provides a new method for treating corneal injury.

The invention provides application of human multifunctional proteoglycan in preparation of drugs for promoting proliferation and / or migration and / or dryness maintenance of corneal limbal stem cells. The human multifunctional proteoglycan has positive effects on proliferation, migration and dryness maintenance of corneal limbal stem cells, is used for preparing the eye drops for treating corneal epithelium injury, can effectively treat corneal limbal stem cell deficiency, promotes corneal epithelium regeneration, and effectively repairs corneal epithelium injury.

The invention provides a method for transdifferentiating corneal limbal stem cells purified in vitro into crystalline lens bodies, and particularly, the method comprises the following steps: inoculating the corneal limbal stem cells onto a culture plate containing a Matrigel glue layer, and culturing the corneal limbal stem cells in a culture system of an LSCM culture solution containing a Y-27632 additive, so that crystalline lens body organs can be obtained. According to the method disclosed by the invention, the corneal limbus stem cells can be quickly transdifferentiated into crystalline lens bodies, so that the corneal limbus stem cells can be implanted into a capsular bag to serve as seed cells for in-situ regeneration of crystalline lenses.

The invention provides application of vasoactive intestinal peptide VIP in preparation of a medicine for preventing and / or treating corneal epithelium lesion, and belongs to the technical field of biological medicine preparation. The invention discloses application of vasoactive intestinal peptide VIP in preparation of a medicine for preventing and / or treating corneal epithelium lesion. Researches show that diabetes can cause expression down-regulation of VIP and a receptor VIPR1 thereof in corneal limbus tissues, so that proliferation activity of corneal limbus stem cells of diabetic mice is reduced, and healing of corneal epithelium wounds is delayed. The exogenous supplement of VIP can effectively reverse the pathological change, significantly enhance the multiplication capacity of corneal limbal stem cells and accelerate the repair of corneal epithelium wounds; on the contrary, the VIPR1 receptor is knocked out, so that the proliferation activity of the corneal limbal stem cells is reduced, and the epithelium repair is delayed. Based on the results, the invention provides a new application of VIP in preparation of medicines for preventing and / or treating corneal epithelium lesions, and a new effective means is provided for repair treatment of the diseases.