37results about How to "Promote in vitro proliferation" patented technology

The invention relates to a novel double-layered composite transmitting tissue regeneration membrane and a preparation method thereof. The double-layered composite transmitting tissue regeneration membrane consists of a PLGA / hydroxyapatite nonporous dense membrane on the outer layer and a PLGA / woolen keratin electrostatic spinning porous membrane on the inner layer. By means of a solvent casting method and a high-voltage electrostatic spinning method, the synthetic material PLGA is combined with the natural component woolen keratin and the inorganic component hydroxyapatite to form the composite material which supplement each other with respective advantages, thereby forming the PLGA / hydroxyapatite / woolen keratin double-layered composite transmitting tissue regeneration membrane. The double-layered composite transmitting tissue regeneration membrane disclosed by the invention is simple in preparation method and moderate in membrane forming conditions, has a good cell biocompatibility, conforms to the requirement of application in the living organism, and has a good application prospect as a novel transmitting tissue regeneration membrane.

The invention relates to a preparation method of a specific extracellular matrix ECM of periodontium. The preparation method comprises the following steps: 1) separating and culturing a human periodontal ligament cell; and 2) preparing the specific extracellular matrix ECM of periodontium; applying the specific extracellular matrix ECM of periodontium and applying the specific extracellular matrix ECM of periodontium as a human periodontal ligament stem cell in in-vitro amplification; and promoting directional differentiation of the ECM. The preparation method is simple and can remarkably promote in-vitro amplification and directional differentiation of the periodontal ligament stem cell.

The invention relates to a novel dual-layer composite bone tissue engineering scaffold and a preparation method thereof. The dual-layer bone tissue engineering scaffold comprises an outer-layer bacterial cellulose membrane and an inner-layer PLGA / MWNTs electrostatic spinning porous scaffold. The synthetic material PLGA, a natural ingredient bacterial cellulose and an inorganic ingredient multi-wall carbon nanotubes are compounded together to form the composite material by using a vacuum freeze drying process and a high-pressure electrostatic spinning method. The materials have complementary advantages, and an ideal dual-layer composite bone tissue engineering scaffold of PLGA / MWNTs / bacterial cellulose, which is excellent in performance and relatively cheap, is built. The dual-layer composite bone tissue engineering scaffold of the PLGA / MWNTs / bacterial cellulose provided by the invention is simple in preparation method, and mild in condition, has good cell biocompatibility, accords with the requirements of application inside a living body, and has a good application prospect as a novel bone tissue engineering scaffold.

The invention discloses a novel culture method for in vitro differentiation from human embryonic stem cells to functional myocardial cells. The novel culture method is different from a conventional typical hanging-drop culture method. According to the novel culture method, an embryoid body is formed by adopting a simplified suspension differentiation culture method, serum-free culture liquid is adopted for improvement, and various inducing and nutrition-enriching substances are added into the culture liquid to increase the differentiation rate of the myocardial cells and prolong the survival time of the myocardial cells; a method for differentiation from embryonic stem cells to myocardial cells is further effectively updated and perfected, the proportion of the myocardial cells differentiated by the embryoid body is greatly increased, and a large quantity of myocardial cell sources with powerful functions can be provided for stem cell clinical transplantation treatment technology, drug screening and the like; the method is simple and feasible to carry out, the culture time is shortened, the functions for in vitro differentiation from the embryonic stem cells to the myocardial cells are improved and include pulsation time as well as autorhythmicity and rhythmicity of myocardial cell beating, and the method is good in repeatability.

This invention discloses a new method of myeloid stem cell external expansion. This invention adjusts the external myeloid stem cells through proper low oxide intensity, and maintains these cells with high hyperplasia state. The method of this invention adopts a controllable and non-trauma physical means and externally adjusts the hyperplasia of myeloid stem cells. It realizes the small amount of sampling, external expansion and large amount of obtaining in order to apply myeloid stem cells to cure many kinds of clinic trauma diseases, retrograde diseases, hereditary defects and etc, and it has good economic and social benefits.

The invention provides a serum-free culture medium for mesenchymal stem cells. The serum-free culture medium specifically comprises a lipid mixture, glutamine, hydrocortisone, zinc sulfate, sodium butyrate, ferric citrate, vitamin C, recombinant epidermal growth factors, recombinant alkaline fibroblast growth factors, RGD oligopeptides, palmitoyl tripeptide-5, palmitoyl tetrapeptide-7, lithium chloride, L-glutathione, a soybean pancreatin inhibitor, beta-mercaptoethanol, ethanolamine, sodium selenite, Hepes, sodium bicarbonate and a basic culture medium. The serum-free culture medium is free of human and animal resources, clear in chemical component, rich in nutrition, capable of achieving UC-MSCs rapid proliferation, and capable of solving the problem of cells number insufficiency and maintaining the biological features and immunophenotyping stability of the mesenchymal stem cells.

The invention relates to the field of protein, in particular to a bioactive peptide GLNMCRQCF as well as a preparation method and application thereof. Amino acid sequence of the bioactive peptide GLNMCRQCF is Gly-Leu-Asn-Met-Cys-Arg-Gln-Cys-Phe. An in-vitro immunomodulatory function experiment verifies that the bioactive peptide GLNMCRQCF has a very good immunomodulatory function. The bioactive peptide GLNMCRQCF can remarkably promote in-vitro proliferation of lymphocytes, improve the immunity of an organism, reduce the incidence rate of the organism, promote activation of macrophages, release cytokines and improve the quality of life, and has very important significance on the development of food, health care products and medicines with immunomodulatory functions.

The invention discloses a method for in-vitro culture of NK cells. It is found that chebulinic acid can promote in-vitro proliferation of the NK cells and can remarkably improve the broad-spectrum killing activity of the NK cells and the killability of cancer cells, that is, the expression of the killing activity biomarker CD107a is significantly improved and the killability of MKN-45 human gastric carcinoma cells is remarkable, and when chebulinic acid intervenes with culture, the phenotype and the content of the NK cells cannot be changed. Therefore, the NK cells can be cultured in vitro byusing chebulinic acid, NK cell proliferation is promoted, the killability of the NK cells is improved, and the chebulinic acid can be added into a culture medium to be developed into a culture mediumspecial for in-vitro culture of the NK cells.

The invention discloses an application of a CXC chemokine receptor 4 gene activator in promoting in-vitro proliferation of mesenchymal stem cells. It is found that cyclo-(4-hydroxy-Pro-Leu)dipeptide has an effect of promoting in-vitro proliferation of bone marrow mesenchymal stem cells, and after intervention culture of the cyclo-(4-hydroxy-Pro-Leu)dipeptide, the bone marrow mesenchymal stem cellsstill have multidirectional differentiation capacity and can be used for seed cells in stem cell engineering. The cyclo-(4-hydroxy-Pro-Leu)dipeptide can exert the effects by activating expression ofCXCR4 in the bone marrow mesenchymal stem cells, and thus the cyclo-(4-hydroxy-Pro-Leu)dipeptide is an effective CXCR4 activator.

The invention discloses a rehmannia extract, a preparation method and application in promoting CIK cell in-vitro proliferation. The total content of rehmannin B, rehmannin D and jioglutin A in the extract is not lower than 90%. The preparation method includes the steps that S1, fresh rehmannia slices are subjected to heat reflux extraction by using an ethanol aqueous solution and subjected to vacuum concentration to obtain an extract; S2, the extract is dissolved with water, extracted with petroleum ether to remove impurities and then extracted with dichloromethane, and dichloromethane extraction solutions are combined, concentrated and dried to obtain a dichloromethane extract; S3, the dichloromethane extract is loaded on an XDA-1B macroporous adsorption resin column, eluted with 30% ethanol to remove impurities and then eluted with 75% ethanol, and 8-10 BV of eluent is collected, concentrated and dried to obtain a crude product of therehmannia extract; S4, the crude product of therehmannia extract is loaded to a normal-phase silica column and eluted with a dichloromethane / methanol / isopropanol mixed solvent in the volume ratio of 10:1:0.2, and 6-7 BV eluent is collected, concentrated and dried. The extract can promote CIK cell in-vitro proliferation.

The invention discloses a purpose of asiatic acid derivatives for promoting in vitro proliferation of human ADSCs (Adipose-Derived Stem Cells) and maintaining stemness. Through studies, people discover that both similarities and differences exist on the effects of asiatic acid, 2,3-dicarbonyl-23-carboxyl n-butyl-ursane-12-ethenyl-28-oxocyclopentanecarboxylate and 2,3-dicarbonyl-23-carboxyl iso-butyl-ursane-12-ethenyl-28-oxocyclopentanecarboxylate on the human ADSCs; each of the asiatic acid, the 2,3-dicarbonyl-23-carboxyl n-butyl-ursane-12-ethenyl-28-oxocyclopentanecarboxylate and the 2,3-dicarbonyl-23-carboxyl iso-butyl-ursane-12-ethenyl-28-oxocyclopentanecarboxylate can obviously promote the in vitro proliferation of the human ADSCs, but the asiatic acid can also promote the chondrogenicdifferentiation of the human ADSCs while promoting the in vitro proliferation of the human ADSCs, and is suitable for an aspect of chondrogenic application of the human ADSCs; and the 2,3-dicarbonyl-23-carboxyl n-butyl-ursane-12-ethenyl-28-oxocyclopentanecarboxylate and the 2,3-dicarbonyl-23-carboxyl iso-butyl-ursane-12-ethenyl-28-oxocyclopentanecarboxylate can maintain the stem cell characteristics of the human ADSCs while promoting the in vitro proliferation of the human ADSCs, and is suitable for the in vitro fast proliferation of seed cells of the human ADSCs.

The invention provides a 3D suspension culture method for in-vitro amplification of spermatogonial stem cells. The method comprises the following steps: inoculating a spermatogonial stem cell suspension on RGD polypeptide modified porous polylactic acid microspheres in a rotary biological reaction container, adding an improved RPMI-1640 culture medium, carrying out three-dimensional static cultureat 30-34 DEG C, starting a rotary system after the spermatogonial stem cells adhere to the RGD polypeptide modified porous polylactic acid microspheres, and carrying out three-dimensional dynamic suspension culture at 30-34 DEG C. By using the method disclosed by the invention, in-vitro large-scale amplification of the mouse SSCs can be realized on the premise of not adding feeder layer cells, and good stem cell proliferation activity and undifferentiated activity are obtained.

The invention discloses an in-vitro amplification method of tumor infiltration lymphocyte (TIL). The method includes the steps of S1, separating the TIL, to be more specific, soaking cut-up para-carcinoma tissue into a collagenase solution for digestion, filtering, performing PBS washing, collecting filtrate, centrifuging, abandoning supernate, adding PBS to prepare a cell suspension, using lymphocyte separation liquid to perform discontinuous density gradient centrifuging, collecting the lymphocyte of the lower interface after the centrifuging is finished, performing PBS washing, culturing ina culture medium containing 10% human AB type serum and IL-2 in a suspending manner according to the concentration of 1*10<6> / mL, and changing liquid once every other 3-5 days; S2, culturing and amplifying, to be more specific, when the lymphocyte in holes grows to 1*10<7>, transferring into a culture medium containing anti-CD3 monoclonal antibody, anti-CD28 monoclonal antibody and medicine for incubation, adding IL-2 after 24 hours to perform culture, and changing a fresh culture medium containing IL-2 with the same concentration every other 2-3 days in a half-amount manner to perform culture for 22-25 days.

The invention belongs to the technical field of sow breeding, and particularly relates to a composite additive bag for promoting mammary gland development of replacement gilts and application of the composite additive bag. The composite additive bag is prepared from an additive and a carrier; the additive is composed of cellulose, inulin, silymarin, baicalin and nicotinic acid; and the carrier iscomposed of zeolite powder and defatted rice bran. The composite additive bag for promoting the mammary gland development of the replacement gilts is researched and prepared according to an apoptosismechanism of mammary gland cells of the replacement gilts, physiological characteristics and reproductive physiological characteristics of mammary gland development of the replacement gilts and physicochemical characteristics of nutrient substances eaten by the replacement gilts. It is found that the weight of mammary glands of the replacement gilts in a third estrus period can be increased, apoptosis of the mammary gland cells in puberty is inhibited, development of mammary gland parenchyma is promoted, then lactation capacity of the gilts in a lactation stage is improved, and productivity and economic benefits of a farm are improved.

The invention discloses an activation method for stem cells based on a colony stimulating factor 1. The CSF1 is used for mobilizing, proliferating and activating the stem cells, and the CSF1 can promote in-vitro proliferation of the stem cells and promote mobilization, proliferation and activation of the stem cells in the body, and is expected to play an important role in cell culture, disease treatment and anti-aging healthcare.

The invention discloses a polypeptide and application thereof in promoting proliferation of human natural killer cells and preparing a human natural killer cell culture medium. The natural killer cell is an anti-tumor and anti-virus innate immune cell, plays an important role in natural immunity, also plays an important role in the aspects of early-stage anti-tumor and immune surveillance, and can directly kill tumor cells without antigen specific recognition. As the number of natural killer cells in a human body is small, large-scale research experiments are limited. The polypeptide has the effect of promoting proliferation of the human natural killer cells and can be used for developing and preparing a culture medium for promoting in-vitro proliferation of the human natural killer cells.

The invention discloses application of rehmannin in promoting in-vitro proliferation of CIK (cytokine-induced killer) cells. The application has the advantages that rehmannin B, rehmannin D and jioglutin A can obviously improve the in-vitro proliferation function of the CIK cells, but jioglutin B does not have the in-vitro proliferation function; compared with the jioglutin A, only the configuration of Cl-connected carbon atoms in jioglutin B is different, and the configurations of carbon atoms in the rehmannin B and the rehmannin D are the same with the configuration of carbon atoms in the jioglutin A, so that the configuration of carbon atoms is critical to the promotion of cell proliferation activity of the CIK cells of the rehmannin type compounds; accordingly, the rehmannin B, the rehmannin D and the jioglutin A can be used for preparing a culture medium for promoting the in-vitro proliferation of the CIK cells, for example, the culture medium can be prepared by adding the rehmannin B, the rehmannin D or the jioglutin A into a RPMI1640 culture solution.

The invention discloses a rehmannia extract, a preparation method and application in promoting CIK cell in-vitro proliferation. The total content of rehmannin B, rehmannin D and jioglutin A in the extract is not lower than 90%. The preparation method includes the steps that S1, fresh rehmannia slices are subjected to heat reflux extraction by using an ethanol aqueous solution and subjected to vacuum concentration to obtain an extract; S2, the extract is dissolved with water, extracted with petroleum ether to remove impurities and then extracted with dichloromethane, and dichloromethane extraction solutions are combined, concentrated and dried to obtain a dichloromethane extract; S3, the dichloromethane extract is loaded on an XDA-1B macroporous adsorption resin column, eluted with 30% ethanol to remove impurities and then eluted with 75% ethanol, and 8-10 BV of eluent is collected, concentrated and dried to obtain a crude product of therehmannia extract; S4, the crude product of therehmannia extract is loaded to a normal-phase silica column and eluted with a dichloromethane / methanol / isopropanol mixed solvent in the volume ratio of 10:1:0.2, and 6-7 BV eluent is collected, concentrated and dried. The extract can promote CIK cell in-vitro proliferation.

The invention discloses an application of asiatic acid to promotion of in vitro proliferation of human adipose mesenchymal stem cells and induction of chondrogenic differentiation of the human adiposemesenchymal stem cells. Researches discover that the effects of the asiatic acid, 2,3-dicarbonyl-23-carboxy n-butyl-ursane-12-ene-28-ethyl carboxylate and 2,3-dicarbonyl-23-carboxy isobutyl-ursane-12-ene-28-ethyl carboxylate on the human adipose mesenchymal stem cells have commonality and difference; the asiatic acid, the 2,3-dicarbonyl-23-carboxy n-butyl-ursane-12-ene-28-ethyl carboxylate and the 2,3-dicarbonyl-23-carboxy isobutyl-ursane-12-ene-28-ethyl carboxylate can significantly promote the in-vitro proliferation of the human adipose mesenchymal stem cells, but the asiatic acid can promote the chondrogenic differentiation of the human adipose mesenchymal stem cells while promoting the in-vitro proliferation of the human adipose mesenchymal stem cells, and is suitable for chondrogenesis of the human adipose mesenchymal stem cells, and the 2,3-dicarbonyl-23-carboxy n-butyl-ursane-12-ene-28-ethyl carboxylate and the 2,3-dicarbonyl-23-carboxy isobutyl-ursane-12-ene-28-ethyl carboxylate can maintain the stem cell characteristics of the human adipose mesenchymal stem cells while promoting the in vitro proliferation of the human adipose mesenchymal stem cells, and are suitable for rapid in vitro proliferation of the human adipose mesenchymal stem cells as seed cells.

The invention discloses a separation, screening, culture and functional identification method of human aortic vascular wall stem cells expressing c-Kit and provides a kit for obtaining human aortic vascular wall stem cells. The kit contains a substance bound with c-Kit positive cells. In the kit, the substance bound with the c-Kit positive cells includes a c-Kit antibody, and / or the c-Kit antibodyis specifically a c-Kit immunomagnetic bead antibody. Tunica adventitia vasorum is separated from human aortic tissue and is digested with collagenase, a cell suspension is obtained through filtration of a cell strainer, c-Kit positive vascular wall stem cells are obtained through screening of the c-Kit immunomagnetic bead antibody, the stem cells have good in-vitro proliferation and transfer ability, has the differentiation potential of mesenchymal stem cells (can be differentiated into fat cells, osteoblasts and cartilage cells) and also has the tissue specificity differentiation capacity of differentiating vascular wall stem cells into vascular smooth muscle cells, and apoptosis occurs rarely.

The invention discloses a method for in-vitro culture of NK cells. It is found that chebulinic acid can promote in-vitro proliferation of the NK cells and can remarkably improve the broad-spectrum killing activity of the NK cells and the killability of cancer cells, that is, the expression of the killing activity biomarker CD107a is significantly improved and the killability of MKN-45 human gastric carcinoma cells is remarkable, and when chebulinic acid intervenes with culture, the phenotype and the content of the NK cells cannot be changed. Therefore, the NK cells can be cultured in vitro byusing chebulinic acid, NK cell proliferation is promoted, the killability of the NK cells is improved, and the chebulinic acid can be added into a culture medium to be developed into a culture mediumspecial for in-vitro culture of the NK cells.

The invention belongs to the field of biological medicine, and relates to a cell membrane for rapid amplification of NK cells and application thereof. The invention also relates to sorting and amplification processes of the NK cells. The invention discloses the cell membrane. 41BBL and IL15 are expressed on the surface of the cell membrane. The invention further relates to a preparation method ofthe cell membrane. According to the invention, a set of NK cell separation and rapid efficient amplification method is established, PBMCs (or the NK cells separated by magnetic beads) are used as an original material, the NK cells are specifically amplified in vitro through stimulation of specific cytokines, and the NK cells are obtained and used for treating malignant tumors.

The invention relates to a novel double-layered composite transmitting tissue regeneration membrane and a preparation method thereof. The double-layered composite transmitting tissue regeneration membrane consists of a PLGA / hydroxyapatite nonporous dense membrane on the outer layer and a PLGA / woolen keratin electrostatic spinning porous membrane on the inner layer. By means of a solvent casting method and a high-voltage electrostatic spinning method, the synthetic material PLGA is combined with the natural component woolen keratin and the inorganic component hydroxyapatite to form the composite material which supplement each other with respective advantages, thereby forming the PLGA / hydroxyapatite / woolen keratin double-layered composite transmitting tissue regeneration membrane. The double-layered composite transmitting tissue regeneration membrane disclosed by the invention is simple in preparation method and moderate in membrane forming conditions, has a good cell biocompatibility, conforms to the requirement of application in the living organism, and has a good application prospect as a novel transmitting tissue regeneration membrane.

The invention discloses application of rehmannin in promoting in-vitro proliferation of CIK (cytokine-induced killer) cells. The application has the advantages that rehmannin B, rehmannin D and jioglutin A can obviously improve the in-vitro proliferation function of the CIK cells, but jioglutin B does not have the in-vitro proliferation function; compared with the jioglutin A, only the configuration of Cl-connected carbon atoms in jioglutin B is different, and the configurations of carbon atoms in the rehmannin B and the rehmannin D are the same with the configuration of carbon atoms in the jioglutin A, so that the configuration of carbon atoms is critical to the promotion of cell proliferation activity of the CIK cells of the rehmannin type compounds; accordingly, the rehmannin B, the rehmannin D and the jioglutin A can be used for preparing a culture medium for promoting the in-vitro proliferation of the CIK cells, for example, the culture medium can be prepared by adding the rehmannin B, the rehmannin D or the jioglutin A into a RPMI1640 culture solution.

The invention discloses peptides and a method for amplifying CIK cells in vitro. Research results of the embodiment of the invention show that a peptide A and a peptide B can effectively promote in-vitro proliferation of CIK cells; meanwhile, technicians in the field know that CD3+CD56+cells are main effector cells of the CIK cells, and the proportion of the CD3+CD56+cells in the cells subjected to induced proliferation of the peptide A and the peptide B is remarkably higher than that of a conventional induced culture method without the peptide A or the peptide B. The high proportion of the CD3+CD56+cells determines that the same number of peptide A and peptide B group cells has stronger tumor lethality in nude mice. Therefore, the peptide A and the peptide B can be used for CIK cell in-vitro amplification to prepare a CIK cell in-vitro amplification reagent.

The invention relates to a method for differentiating mesenchymal stem cells into bile duct cells. The method comprises the following steps: (1) differentiating human mesenchymal stem cells into endodermal stem cells (cell differentiation culture; and (2) carrying out maturation differentiation on the endodermal stem cells, so as to obtain the bile duct cell (cell maturation culture). According toa ''two-step differentiation method'' for inducing the differentiation of the mesenchymal stem cells under a specific culture condition, the human mesenchymal stem cells is successfully differentiated into the functional bile duct cells, the stem cells and other cells do not need to be subjected to co-culture or genetic recombination, and the induced differentiation is carried out only based on the specific culture condition. The bile duct cells are easily available, have no immunoreaction and tumorigenesis property, can be rapidly proliferated in vitro and are hopeful of being applied to clinical drug screening, manual biliary tract reconstruction and the like.

The invention discloses application of caffeic acid derivatives to promotion of in-vitro proliferation of umbilical cord mesenchymal stem cells and preparation of a proliferation promoting culture medium. By experiments in the invention, the following result can be found that caffeic acid-4-2-hydroxyethyl morpholine ester can obviously promote in-vitro proliferation of hUCMSCs (human umbilical cord mesenchymal stem cells), has a dose-dependent effect and a time-dependent effect, and the hUCMSCs proliferated by the caffeic acid-4-2-hydroxyethyl morpholine ester are still kept having very high multi-directional differentiation capacity and cna be used as seed cells. Therefore, the caffeic acid-4-2-hydroxyethyl morpholine ester can be used for promoting in-vitro proliferation of the hUCMSCs and used for preparing a culture medium for promoting in-vitro proliferation of the hUCMSCs.