179 results about "Primary culture" patented technology

The invention relates to a marine ranching digital construction method and system based on artificial intelligence, and the method comprises the steps: monitoring plankton in a marine ranching in real time, obtaining a water body image through an image recognition technology, and building a dynamic database, so as to regularly store and update the related data of the plankton. Based on monitoring data, a water quality evaluation model is established, and the correlation among plankton species, density and water quality parameters is analyzed. Factors such as water flow velocity and flow direction are integrated, a flow change model is established, and the influence of water flow on plankton is judged. A primary culture optimization scheme is formulated by simulating the interaction between cultured organisms and planktons, and the culture trend is predicted in real time and the management strategy is adjusted by integrating the water quality evaluation and the output result of the flow change model. According to the scheme, the intelligent management level of the marine ranching is effectively improved, dynamic and scientific management of the culture environment is achieved, powerful support is provided for healthy growth of cultured organisms, and sustainable development of the marine ranching is promoted.

The invention provides application of a bacteriostatic agent in open type tissue culture and rapid propagation of polyploidy bamboo reeds, and relates to the technical field of tissue culture and rapid propagation of plants, the bacteriostatic agent is applied to the open type tissue culture and rapid propagation process of the polyploidy bamboo reeds, and the use method is that the bacteriostatic agent is added into a culture medium for use or sprayed or oscillated and flushed for use, and the bacteriostatic agent is applied to the open type tissue culture and rapid propagation process of the polyploidy bamboo reeds. The bacteriostatic agent is formed by mixing isothiazolinone and sodium dehydroacetate, the mass addition ratio of the isothiazolinone to the sodium dehydroacetate is (8-5): (2-5), and the method comprises the following steps: S1, primary culture of explants; s2, multiplication culture of seedlings; s3, rooting culture, wherein each liter of the culture medium contains 0.05-1g of the bacteriostatic agent in the above steps. The method has the beneficial effects that under the condition that the method does not need to be controlled in a sterile environment and does not depend on the quality of an inoculator, a large amount of propagation of the polyploidy bamboo reed tissue culture seedlings is realized by adopting an open tissue culture technical mode, the productivity speed of a tissue culture center is improved, and the construction threshold and cost of the tissue culture center are reduced.

The invention provides a method for inducing test tube tuberous roots under dahlia pinnata tissue culture conditions, which belongs to the technical field of plant tissue culture and comprises the following steps: step 1, obtaining sterile explants; step 2, primary culture: removing a color-changing part on the wound surface of the to-be-inoculated explant obtained in the step 1, and then inoculating the to-be-inoculated explant into a primary culture medium for culture to obtain a sterile test-tube plantlet; step 3, subculture multiplication culture: cutting the seedlings into cluster blocks, and inoculating the cluster blocks into a subculture multiplication culture medium to obtain multiplication test-tube seedlings; step 4, rooting culture; performing illumination culture in a rooting culture medium for 4 weeks to obtain rooted test-tube plantlets; 5, test tube root tuber induction: inoculating the rooted test tube plantlet into a root tuber induction culture medium for illumination culture for 4 weeks to obtain dahlia pinnata test tube sterile root tubers; through segmented induction and addition of different exogenous hormones, the breeding efficiency of dahlia pinnata can be effectively improved, and the breeding period is shortened.

The invention relates to an extraction and culture method of urine-derived stem cells, and belongs to the field of cell biology and regenerative medicine. The method comprises the following steps: coating a culture plate with a gelatin solution; collecting clean midstream urine of healthy people and adding antibiotics; centrifugally separating the cell precipitate; washing with a phosphate buffer solution; re-suspending the precipitate by using a urine stem cell complete culture medium for primary culture; and after the cells grow to a certain fusion degree, carrying out passage amplification. The urine stem cell complete medium comprises a basic medium, fetal calf serum, epidermal growth factors, platelet-derived growth factors and basic fibroblast growth factors. By adopting the method disclosed by the invention, the urine-derived stem cells can be efficiently and stably separated from urine, and the obtained cells have high multiplication capacity, good stemness maintaining capacity, multidirectional differentiation potential and immunoregulation function, and can provide an ideal seed cell source for treating diseases such as diabetic erectile dysfunction.

The invention relates to the technical field of aquatic animal cell culture, in particular to a rapid enrichment and separation method for gonad somatic cells of plectropomus leopardus. Comprising the following steps: selecting healthy plectropomus leopardus for anesthesia; the method comprises the following steps: taking gonad tissues, performing sterile treatment, cutting, centrifugal washing and filtering, inoculating the gonad tissues into a culture medium for primary culture to obtain gonad cells emigrated from tissue blocks, performing trypsin-EDTA digestion and centrifugal washing, and treating the gonad cells with 1 * PBS containing polysorbate; after closing, antibody incubation and washing, the marking effect is verified through a fluorescence microscope, and specific enrichment of gonad somatic cells is achieved; and filtering and purifying the enriched fluorescence-labeled cells, placing the cells under a fluorescence microscope, accurately picking fluorescence-positive cells under a fluorescence visual field, transferring the fluorescence-positive cells into a culture dish containing a complete medium, inoculating the separated fluorescence-positive cells into the optimized complete medium, and carrying out purification culture, so that the cells grow in a wall-attached manner and stable passage is realized.

The application discloses a construction method and application of an ovary cell line of Quasipaa spinosa. The method comprises the following steps: selecting metamorphosis period larva ovary tissue, disinfecting and obtaining the ovary tissue, and then using collagenase I digestion solution to digest the ovary tissue at 26 DEG C for 1 hour; after the tissue block adheres to the wall, using a low-osmotic-pressure complete culture medium to carry out primary culture at 25-27 DEG C, wherein the culture medium is 60% diluted DMEM / F12 culture medium containing 10%-15% fetal bovine serum; when the cell confluence degree is higher than 70%, using trypsin-EDTA to digest the cells for 1-2 minutes to subculture, and the subculture culture medium contains 30% cell conditional culture medium; and finally, the cells are cryopreserved through programmed cooling. The ovary epithelial cell line of Quasipaa spinosa is successfully established for the first time, the survival rate of the cells after recovery is high, and the karyotype is stable. The constructed cell line can be used for amphibian reproductive biology research, compound screening and in-vitro conservation of germplasm resources.

The invention discloses a culture medium composition and a culture method for in-vitro rapid propagation culture of hippeastrum hippeastrum, and belongs to the technical field of plant tissue culture. The adenine, the rhizoma polygonati and the wormwood extract are added into the hippeastrum striatum in-vitro rapid propagation culture medium for the first time, and the culture medium combination for hippeastrum striatum in-vitro rapid propagation culture is developed. The hippeastrum hippeastrum in-vitro rapid propagation culture method established on the basis of the culture medium combination can be used for in-vitro rapid propagation culture of different varieties of hippeastrum hippeastrum, the germination rate of primary culture in the hippeastrum hippeastrum in-vitro rapid propagation culture can be increased, and the differentiation coefficient and the seedling formation coefficient in the hippeastrum hippeastrum tissue culture process can be increased. The efficient and stable hippeastrum striatum in-vitro rapid propagation method established by the invention lays a technical foundation for accelerating a hippeastrum striatum breeding process and commercialized production.

The invention belongs to the technical field of sweet potato seedling detoxification, and particularly relates to a sweet potato seedling detoxification rapid propagation method and application, and the sweet potato seedling detoxification rapid propagation method comprises the following steps: carrying out heat treatment culture on sweet potato seedlings at 36-38 DEG C, disinfecting, and taking stem segments with the length of 0.5-1.5 cm as explants; inoculating the explant into a primary culture medium for culturing; after multiple shoots are induced from the explants, inoculating the explants into a subculture medium for culturing; cutting the cultured cluster buds into single buds, inoculating the single buds into a rooting culture medium for culturing, and detecting the detoxification effect after bud seedlings root to obtain detoxified sweet potato tissue culture seedlings. By means of the method, virus detoxification of the sweet potato seedlings can be achieved, the detoxification rate reaches up to 95%, the survival rate reaches 96.7%, the seedling breeding period of 2-3 months is shortened to 45-50 days, the health level and breeding efficiency of the seedlings are improved, and the yield and quality of sweet potatoes are further improved.

The application discloses a culture medium for tissue culture of Limosella australis, which comprises a primary culture medium, a proliferation and differentiation culture medium and a rooting culture medium. The primary culture medium is MS+6BA 0.4-0.6 mg / L+NAA 0.04-0.06 mg / L+AC 0.8-1.2 mg / L+2,4-D 0.04-0.06 mg / L. The proliferation and differentiation culture medium is modified MS+6BA 0.09-0.11 mg / L+NAA 0.04-0.06 mg / L+AC 1.5-2.5 mg / L. The rooting culture medium is 1 / 2MS+IAA 0.09-0.11 mg / L+AC 2.5-3.5 mg / L. The technical scheme has the highest callus induction rate of 86.7%, the highest differentiation rate of 86.7% and the highest rooting rate of 93.3%.

The invention relates to the technical field of seedling raising, and discloses an efficient tissue culture and rapid propagation seedling raising method for xanthoceras sorbifolia bunge, which comprises the following steps: selecting and pretreating explants, selecting current-year robust branches of the xanthoceras sorbifolia bunge, cutting stem segments with axillary buds as the explants, cleaning, trimming and disinfecting the explants, performing surface disinfection on the pretreated explants, and performing tissue culture and rapid propagation seedling raising on the pretreated explants to obtain the xanthoceras sorbifolia bunge. When the Xanthoceras sorbifolia Bunge is subjected to efficient tissue culture and rapid propagation seedling culture, an operation standard considering thorough sterilization and tissue activity protection is established by standardizing an explant selection standard and executing a step-by-step surface disinfection procedure, so that the microbial contamination rate caused by incomplete disinfection of the explant can be reduced, meanwhile, tissue activity damage caused by excessive disinfection is reduced, and the survival rate of the Xanthoceras sorbifolia Bunge is improved. Therefore, sufficient and healthy sterile initial materials are guaranteed to be obtained, the problems of high primary culture pollution rate and induction starting failure are solved, and a reliable foundation is laid for establishment of a whole tissue culture and rapid propagation system.

The invention relates to an isolated culture method of functional Chinese softshell turtle ovarian granular cells. The isolated culture method comprises the following steps: S1, collecting ovarian tissues: collecting Chinese softshell turtle ovarian tissues; s2, separating and collecting a granular cell layer; s3, digesting and collecting granular cells; s4, primary culture of granulosa cells: transferring the collected cells into a cell culture bottle containing 4 ml of a complete culture medium for primary culture, wherein the complete culture medium comprises the following main components: a DMEM culture medium, 15% of fetal calf serum, 1% of penicillin streptomycin and 1% of an insulin-transferrin-selenium additive; and S5, subculturing the granular cells. The isolated culture method and the culture medium system provided by the invention are simpler and more time-saving and labor-saving; according to the method, the aromatase CYP19A1 can be expressed normally, the characteristics of the granular cells are kept, the aromatase CYP19A1 can be expressed normally, it is indicated that the in-vitro cultured cells can keep the functionality of the cells, and the in-vitro proliferation capacity of the granular cells is improved to a great extent.

The invention discloses a factory-like efficient breeding method and system for virus-free seed potatoes in a potato bottle, and relates to the technical field of agricultural biology. The method comprises the following steps: selecting healthy buds, stripping stem tips, carrying out primary culture to obtain seedlings, screening virus-free seedlings through RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection, and transferring the virus-free seedlings to a culture flask containing a liquid MS culture medium for expanding propagation; hardening the propagated virus-free seedlings to obtain stress-resistant virus-free seedlings, inoculating the stress-resistant virus-free seedlings into a tissue culture bottle containing an induction culture medium, performing induction culture under set illumination and temperature, and dynamically adjusting a nutrient solution to obtain virus-free seed potatoes; and classifying and screening the detoxified seed potatoes and storing in a specific environment. The system comprises a sterile operation unit, a multi-stage culture unit, a seedling hardening unit and a graded storage unit. The method can efficiently breed the virus-free potato seeds, reduces the virus infection risk, improves the breeding efficiency and the potato seed quality, and is suitable for industrialized production.

The invention discloses a stipa purpurea tissue culture and rapid propagation method and a special culture medium. The method comprises the following steps: (1) selecting and pretreating an explant; (2) primary culture; (3) subculture; (4) rooting culture; and (5) seedling hardening and transplanting. Through an optimized explant disinfection process, a culture medium system based on N6, a specific plant growth regulator combination and addition of enzyme hydrolysis casein in a rooting stage, a breakthrough of a stipa purpurea tissue culture and rapid propagation technology is achieved, the pollution rate is lower than 5%, the multiplication coefficient reaches 4.2 or above, the rooting rate exceeds 93%, the transplanting survival rate reaches up to 89%, and the tissue culture and rapid propagation technology has the advantages that the tissue culture and rapid propagation efficiency is high. The whole rapid propagation period only needs 60-70 days, and efficient and reliable technical support is provided for large-scale and standardized production and ecological restoration application of stipa purpurea.

The invention belongs to the technical field of plant tissue culture, and provides a composite systemic bactericidal solution for lily bulb pretreatment and a matched disinfection and primary culture method thereof. Each liter of the working solution comprises 4-8 g / L of 50% carbendazim wettable powder, 2-6 g / L of 50% chlorobromoisocyanuric acid soluble powder and 1-2 g / L of metalaxyl-hymexazol aqueous solution. The method sequentially comprises the steps of appropriate dehydration of bulbs, soaking in a composite systemic bactericidal solution, standing for absorption, surface sterilization, short soaking in a protective solution and continuous bacteriostatic culture. According to the technical scheme, the limitation of traditional surface sterilization is broken through, deep and surface double sterilization is considered, the multi-mechanism and multi-link synergistic effect is achieved, the pollution rate (smaller than or equal to 3%) of lily bulb primary culture is reduced, mercuric chloride is avoided, and the method is safe, environmentally friendly and suitable for large-scale culture of lilies and other bulb crops.

The invention relates to the technical field of plant tissue culture, in particular to a method for tissue culture and rapid propagation of polygala japonica. Comprising the following steps: sterilizing and disinfecting explants; carrying out primary culture on the sterilized and disinfected explant; carrying out multiplication culture on primary cluster buds; carrying out rooting culture on rootless tissue culture seedlings; domesticating and transplanting rooting tissue culture seedlings. A primary culture medium, a proliferation culture medium and a rooting culture medium are reasonably designed according to different growth regulator formulas by taking a stem section with buds of polygala japonica as an explant material and are respectively used for performing clumpy bud induction, clumpy bud proliferation and rooting induction on the explant material, the total culture process only needs about 100 days, the breeding efficiency is high, and the survival rate is high. The inductivity, the tufted bud multiplication coefficient and the rooting rate of the explant tufted buds all reach a relatively high level, the cultured tissue culture seedlings are good in quality and high in transplanting survival rate, the tissue culture and rapid propagation effects of the polygala japonica are good, and a solid foundation is laid for realizing industrialized production of the polygala japonica.

The invention discloses a siRNA molecule for interfering sheep TBXT gene expression in primary culture of sheep bone marrow mesenchymal stem cells and application of the siRNA molecule, and relates to the technical field of molecules. According to the invention, a method for in-vitro targeting sheep TBXT gene interference siRNA transfected cells is established by synthesizing a specific targeting combination sheep TBXT gene small fragment sequence, and the mRNA expression level of the TBXT gene of sheep bone marrow mesenchymal stem cells can be effectively interfered. A further research proves that the proliferation efficiency of the sheep bone marrow mesenchymal stem cells is obviously reduced after the TBXT gene is interfered. The TBXT gene is helpful for researching the biological function of the TBXT gene participating in embryonic caudal vertebra development regulation, and has good application value for researching the molecular mechanism of the sheep tail type character.

The application discloses a tissue culture propagation method of eriochloa contracta. The creeping stem section of the eriochloa contracta is collected, washed, and then sterilized and dried; the sterilized and treated stem section is inoculated into a bud induction medium of MS+1-1.5 mg / L 6-BA+0.1-0.5 mg / L NAA+30 mg / L sucrose+6.5 mg / L agar to perform bud induction, and after 15-20 days of culture, the adventitious bud seedling is obtained; the adventitious bud of about 1-1.5 cm induced in the primary culture medium is inoculated into a proliferation culture medium to perform subculture, and the next step can be divided into single bud expansion or direct transplanting domestication. The two-step method only needs primary culture-proliferation and rooting to be simultaneously performed, and the transplanting survival rate is high.

Disclosed is a chemically defined airway basal stem cell culture medium. The provided culture medium involves a simple preparation method and high safety, and does not require a feeder layer or a matrigel-coated culture plate. By means of the culture medium, primary cultured airway basal stem cells achieve good stability; and after multiple passages, the normal morphology of the basal stem cells can be maintained and good differentiation potential can be maintained. In addition, the culture medium has clear components, and can meet both scientific research-grade and clinical-grade applications of airway basal stem cells.

The invention discloses an osteoclast culture method as well as a culture medium and application thereof, the culture method is based on primary monocyte extraction, inoculation is performed according to a specific density, then quantitative M-CSF is added, and culture is performed overnight; on the second day, when the cell confluence degree of the mononuclear bone marrow hematopoietic stem cells is 5-30%, supplementing quantitative RANKL into each hole, continuously culturing for 2-4 days, then changing the solution, and obtaining mature osteoclasts on the sixth-seventh day. According to the invention, the optimization relationship among the cell activity, the cell density and the culture medium in the primary culture process of the osteoclast is deeply studied, and from the perspective of cost reduction and efficiency improvement, the use cost of cell factors is remarkably reduced, the number of times of liquid change is reduced, and the traditional culture time of 10-12 days (calculated from cell extraction) is shortened to 6-7 days. The average diameter of the mature osteoclast cultured by the method is larger, the number of fused cell nucleuses is larger, the space proportion of the mature osteoclast is not less than 80%, and the cell quality is obviously higher than that in the prior art.

The application discloses a tissue culture and rapid propagation method for Yangmei of Alocasia macrorrhiza and a special combined culture medium thereof, and the method comprises the steps of sterilizing an explant, carrying out primary culture and subculture and proliferation culture in stages, wherein the primary culture is performed by first inoculating a stem segment into an induction culture medium containing 6-BA 0.8-1.2 mg / L, IBA 0.05-0.15 mg / L and PVP-40 to induce germination, and then transferring the stem segment into an elongation culture medium containing 6-BA 0.1-0.5 mg / L, NAA 0.01-0.05 mg / L and PVP-40 to promote elongation; the new shoots are then transferred into a subculture medium containing 6-BA 1.0-1.8 mg / L, NAA 0.1-0.2 mg / L, GA3 0.5-1.5 mg / L and PVP-40 to proliferate. The application effectively solves the technical problems that Yangmei is prone to brown and death, and it is difficult to simultaneously achieve budding and high growth by means of two-stage hormone stepwise regulation and cooperation with a brown prevention agent, and realizes efficient and large-scale breeding of high-quality seedlings.

The invention belongs to the technical field of in-vitro cultured calculus bovis, and provides in-vitro cultured calculus bovis and a preparation method thereof. The preparation method comprises the following steps: (1) separating cells; (2) primary culture; (3) cell inoculation and cultivation: adding the serum-free culture medium into the carrier matrix, putting the carrier matrix into a reactor, and performing cell inoculation on the carrier matrix after passage; the inoculated culture comprises a proliferation period; a synthesis period; a crystallization period; and (4) collecting and purifying the calculus bovis, namely stopping culturing after 45 days, and sequentially collecting, cleaning, degreasing, drying and crushing the calculus bovis. The preparation method aims at solving the problems of long culture period, unstable content of effective components, high production cost, large batch difference and the like in the existing in-vitro calculus bovis culture technology by optimizing a culture medium formula, regulating and controlling a culture process and a purification process.

The invention discloses a method for separating, culturing and identifying porcine muscle-derived mesenchymal stem cells and fat progenitor cells. The method mainly comprises the following steps: dissociating pig muscle tissues through a tissue processor, obtaining mesenchymal stem cells and adipose progenitor cells by utilizing a flow cytometry sorting technology, and completing in-vitro culture and identification of the cells. According to the technical scheme provided by the invention, the technical problems of primary culture, separation and identification of the mesenchymal stem cells and the adipose progenitor cells of the porcine muscle tissue are effectively solved, and a reliable method is provided for further application of a flow cytometry sorting technology and a myogenic mesenchymal stem cell and adipose progenitor cell primary model in animal nutrition research.

The invention discloses a tissue culture, rapid propagation and rooting culture method for blueberry yunnanensis and application of the tissue culture, rapid propagation and rooting culture method. The culture method disclosed by the invention comprises the following steps of: cleaning and disinfecting an explant of a stem segment of the blueberry yunnanensis, carrying out primary callus culture, carrying out primary callus induction culture, carrying out subculture differentiation culture, carrying out rooting culture and transplanting rooted seedlings. Through disinfection with 15% sodium hypochlorite, the contamination rate can be 0, the germination rate in the callus primary culture process reaches 88.89%, the browning rate in primary callus induction culture is only 13.89%, the browning rate is reduced by more than 30%, the callus yield reaches 96.77%, the callus yield is improved by more than 10%, the callus buds and differentiates in the callus subculture, buds with good growth vigor are formed, and the germination rate of the callus is increased by more than 10%. The rooting rate in rooting culture reaches 80%, and the survival rate after transplanting reaches 83%.

The invention discloses a phantom liquidambar formosana axillary bud tissue culture proliferation method, which takes phantom liquidambar formosana axillary buds as tissue culture materials and improves the propagation speed of phantom liquidambar formosana seedlings. After axillary buds of the 'phantom' liquidambar formosana hance are collected and treated, the most suitable culture medium and growth regulator are screened through primary culture, subculture and rooting culture, so that tissue culture proliferation of the 'phantom' liquidambar formosana hance is completed. According to the method, the population cardinal number of the phantom liquidambar formosana hance can be rapidly increased, the problem that the grafting speed is not high due to the fact that branches of the phantom liquidambar formosana hance are thickened slowly is solved, and a clonal germplasm resource nursery of the phantom liquidambar formosana hance can be built early.

The invention discloses a method for propagating robust artificial seed stems of pinellia ternate through low-cost tissue culture. The method comprises the steps of germination, explant disinfection, establishment of a primary culture system, petiole subculture propagation, tuber subculture propagation and artificial seed stem harvesting. The plant regeneration approach designed by the invention is to induce one end of the petiole to directly generate a small tuber with a complete form under hormone-free concentration, and the small tuber is very similar to the part and the form of the bulbils generated under natural conditions. Tubers are not subjected to a callus stage, are uniform in size and high in hereditary stability, can maintain the characteristics of excellent varieties, and are an optimal way for in-vitro rapid propagation of pinellia ternate. A large number of mature, full, neat and consistent third-generation tubers with high germination rate, stress resistance and storage resistance can be obtained in a short time, the tubers can be directly transplanted to a field without special domestication, the survival rate is high, practical operation and large-scale production are facilitated, and production is not limited by seasons.

The invention relates to the technical field of high-yield collagen MSC (mesenchymal stem cell) engineering construction, and particularly discloses a fixed-point controllable high-yield collagen MSC engineering construction method, which comprises the following steps: 1, MSC separation and primary culture: separating from mammal bone marrow, umbilical cord or adipose tissue to obtain primary MSC, adopting a DMEM / F12 culture medium containing 13% of fetal calf serum, and carrying out primary culture on the primary MSC; carrying out primary culture and passage in a culture environment of 37 DEG C and 5% CO, and screening the fourth generation of MSC in the logarithmic phase as an engineered starting cell; and 2, constructing a collagen synthesis related gene fixed-point editing carrier: designing a fixed-point editing element aiming at a collagen synthesis key gene and an MSC differentiation regulation gene. According to the site-specific controllable high-yield collagen MSC engineering construction method, the collagen expression quantity of the engineered MSC can be greatly improved, the directional synthesis proportion of the type I collagen and the type III collagen is improved, and the requirement of industrialization for high-yield and high-purity collagen is effectively met.

The invention relates to a lung fibroblast culture method, and belongs to the technical field of cell culture. Comprising the following steps: S1, organization preparation; s2, tissue treatment; s3, primary culture: the treated tissue blocks are attached to a culture dish, a culture solution is added, the culture solution is a primary culture system, and culture is performed in an incubator for several days until the culture dish is full of fibroblasts; s4, subculture: digesting the cells subjected to primary culture by adopting conventional 0.25% trypsin, then adding a culture solution which is a subculture system for subculture, and carrying out subculture to a fourth generation; and S5, cell identification. According to the invention, primary and subculture are respectively purified and amplified by virtue of a double culture system of fibroblasts, so that the purification of target cells is facilitated, the cell quality is good, a low-serum growth additive is used as a nutrient substance, the low-serum growth additive is more easily utilized by the cells, the proliferation of the cells is facilitated, and good proliferation activity and phenotypic stability are still kept when the cells are subcultured to the fourth generation; no obvious differentiation phenomenon occurs.

The culture medium comprises a primary culture medium, a proliferation culture medium, a differential culture medium and a rooting culture medium, and the primary culture medium is used for inducing calluses from explants; the proliferation culture medium is used for culturing adventitious buds from the calluses; the differential culture medium is used for culturing true leaves from the adventitious buds; the rooting culture medium is used for enabling true leaves to root; the primary culture medium, the proliferation culture medium, the differential culture medium and the rooting culture medium are sterilized sterile culture mediums; by adopting the culture medium disclosed by the invention, the highest induction rate, the highest proliferation rate, the highest differentiation rate and the highest rooting rate are 95%, and the effect is remarkable.

The present invention provides a method for promoting the differentiation of mesenchymal stem cells (MSCs) into oligodendrocyte progenitor cells, which includes primary culturing the isolated MSCs, followed by secondary passage culturing, then treating them with a gene expression vector containing the "key gene for oligodendrocyte progenitor cell differentiation", and further performing differentiation culture using the "oligodendrocyte progenitor cell differentiation medium". Through specific cell differentiation treatment and culture processes, the present invention uses a cell culture matrix treated with recombinant human laminin to culture MSCs and promote their differentiation, so as to obtain oligodendrocyte progenitor cells derived from MSCs in a relatively short time. The present invention further provides a method for efficiently harvesting oligodendrocyte progenitor cells. Specifically, through a rate-limiting shaking method, oligodendrocyte progenitor cells are harvested and allowed to further proliferate, and a relatively large amount of oligodendrocyte progenitor cells with high purity can be harvested in a short time for the production of cell therapy products.

The invention relates to the technical field of biological culture of algae, in particular to a culture method of Yunnan hot spring algae. Comprising the following steps: inoculating algae species of Yunnan hot spring algae into a culture medium, standing and culturing in a dark place, and filtering to obtain activated algae species; inoculating the activated algae species into a culture bottle, adjusting the pH value to 7.5-8.5, supplementing a sulfur-containing compound, putting the culture bottle on a constant-temperature shaking table, illuminating, culturing for 5-7 days, and filtering to obtain primary culture algae species; inoculating the primary culture algae species into a closed reactor, then adjusting the pH value to 7.5-8.5, carrying out programmed heating and illumination, supplementing a sulfur-containing compound, and culturing for 8-10 days. The temperature control system is used for accurately controlling the temperature of the culture environment, the living environment of the hot spring algae is maintained by monitoring pH and dissolved oxygen in real time, the closed photobioreactor is used for preventing infectious microbe pollution, controllable culture and stable culture of the hot spring algae are achieved, and the yield of the hot spring algae is increased.