75 results about "Fibroblast cultures" patented technology

Methods are provided for classification of cancers by the expression of a set of genes referred to as the core serum response (CSR), or a subset thereof. The expression pattern of the CSR in normal tissues correlates with that seen in quiescent fibroblasts cultured in the absence of serum, while cancer tissues can be classified as having a quiescent or induced CSR signature. Patients with the induced CSR signature have a higher probability of metastasis. Classification according to CSR signature allows optimization of treatment, and determination of whether on whether to proceed with a specific therapy, and how to optimize dose, choice of treatment, and the like.

The invention provides improved methods for producing induced pluripotent stem cells (iPSC) from adult fibroblasts. The methods include contacting adult fibroblasts with a reprogramming composition suitable for reprogramming the adult fibroblasts to iPSC, under conditions effective for the reprogramming composition to penetrate the adult fibroblasts, followed by culturing the contacted fibroblasts for a time period sufficient for the cells to be reprogrammed. The cultured cells are then sorted to select cells based upon their expression of the cell membrane surface markers CD13NEG SSEA4POS Tra-1-60POS. iPSC colonies are then identified from the sorted cells.

An autologous topical formulation containing conditioned medium obtained from culture of autologous fibroblasts has been developed. Unlike other topical formulations, it is autologous since it is derived solely from cells obtained by the person who is to use the formulation. This avoids any possible reaction with proteins derived from the cells. Preferred formulations include gels, creams, lotions, and ointments. The topical formulations of conditioned medium obtained by culturing autologous dermal fibroblasts are topically administered to individuals for the prevention and treatment of scarring, reduction in signs of aging, and improvement in quality of skin.

An autologous topical formulation containing conditioned medium obtained from culture of autologous fibroblasts has been developed. Unlike other topical formulations, it is autologous since it is derived solely from cells obtained by the person who is to use the formulation. This avoids any possible reaction with proteins derived from the cells. Preferred formulations include gels, creams, lotions, and ointments. The topical formulations of conditioned medium obtained by culturing autologous dermal fibroblasts are topically administered to individuals for the prevention and treatment of scarring, reduction in signs of aging, and improvement in quality of skin.

Personalized skin-specific topical formulations containing conditioned medium obtained from cultures of fibroblasts has been developed. Unlike other topical formulations, these formulations are specific for the recipient-skin type specific, and conditioned medium is obtained from a subject with desired skin characteristics. For example, since African American skin is known to wrinkle less and later than Caucasian skin, conditioned medium may be obtained from African American skin for application to pale skin. In another embodiment, the fibroblasts are obtained from young skin for administration to older skin. In still another embodiment, the fibroblasts are obtained from skin that does not suffer from acne or discoloration, for application to skin that is prone to acne or discoloration. Examples of skin donor selection criteria include delayed wrinkling, small pores, resistance to sunburn, resistance to acne, uniform coloration or lack of blotching or age spots and good moisture retention.

Disclosed is a method of inducing a reduced immune response to a transplant in a recipient by treating said recipient with an amount of fibroblasts or a supernatant from a fibroblast culture effective to reduce or inhibit host rejection of the transplant. The fibroblasts or a supernatant from a fibroblast culture can be administered before, at the same time as, or after the transplant. This method is effective in reducing an immune response to a transplant without compromising the immune response to other foreign antigens. Also disclosed is a method of inducing a reduced immune response against a host by foreign tissue, i.e., graft versus host disease.

The invention relates to a method for culturing seed cells of tissue engineering. The method for amplifying the seed cells of the tissue engineering comprises the following steps of: (1) adding a macroporous microcarrier into a bioreactor, hydrating and sterilizing and then adding 50 to 100ml of fibroblast culture medium, and adding fibroblasts for culturing; (2) inhibiting the fibroblasts from amplifying when the fibroblasts are attached to the microcarrier and the growth area is near 50 percent of surface area of the microcarrier, and standing and then pouring out culture solution; and (3) washing the obtained carrier by using sterile, calcium-free and magnesium-free phosphate buffered solution (PBS), then adding an epidermal cell culture medium and epidermal cells, and continuing to stir and culture until the cells overgrow the carrier to obtain the seed cells of skin tissue engineering. The quantity and function of the cells cultured by the method can both be apparently increased,the damage of the cells is reduced, the cells can be directly used as seed cells for skin construction of the tissue engineering, and the effect is superior to those of methods using epidermal cell suspension and epidermal cell patches.

The invention discloses a culture method for fibroblast derived from autologous body skin. The method comprises the following steps: 1) taking autologous body skin, dividing the autologous body skin into multiple tissue blocks, attaching the tissue blocks to a culture dish, dropwise adding concentrated feta calf serum on the tissue blocks, completely soaking the tissue blocks, preventing the tissue blocks from drying and promoting the fibroblast to crawl out; 2) adding a selective culture medium into the culture dish in the step 1), placing the culture dish into an incubator to culture until the fibroblast fusion is finished, wherein the selective culture medium is used for promoting the fibroblast to grow and inhibiting other parenchyma cells to grow; 3) taking the culture dish in the step 2) out of the incubator, adding pancreatin for digestion to enable the fibroblast to fall off from the culture dish; 4) inoculating the fibroblast obtained in the step 3) into a culture bottle by a complete culture medium, enabling the fibroblast density to reach 90 percent to obtain primary fibroblast, wherein the complete culture medium is used for ending the digestion in the step 3) and is used as a culture medium for growth and proliferation of the fibroblast; 5) passaging the bottle-expanding culture of the primary fibroblast in the step 4).

The invention relates to a method for culturing autologous tissue fibroblast, which is characterized by orderly comprising a skin specimen treatment step and a fibroblast culture step, wherein the fibroblast culture step comprises a primary fibroblast culture step; and a fibroblast subculture step, a fibroblast cryopreservation step and a fibroblast unfreezing resuscitation step can be set according to actual needs. The method massively cultures the autologous tissue fibroblast by culture in vitro and amplification of autologous fibroblast; and the autologous tissue fibroblast is used for reinfusion treatment on skin collagen defective disease or degenerative disease simply and conveniently with good safety, and does not generate rejection reaction and has obvious effect when used.

Certain embodiments here in are directed to a method of treating a tissue associated with a defect in a human including wrinkles, rhytids, depressed scar, cutaneous depressions, stretch marks, hyperplasia of the lip, nasolabial fold, melolabial fold, scarring from acne vulgaris, and post-rhinoplasty irregularity. The tissue defect may be treated by introducing a plurality of in vitro cultured autologous fibroblast cells at or proximal to the defect area of the patient's tissue. The autologous fibroblast cells may have been cultured in vitro to expand the number of fibroblast cells in at least one medium that comprises autologous serum. The autologous fibroblast cell cultures may be derived from connective tissue, dermal, fascial fibroblasts, papillary fibroblasts, and / or reticular fibroblasts.

Certain embodiments here in are directed to a method of treating a tissue associated with a defect in a human including wrinkles, rhytids, depressed scar, cutaneous depressions, stretch marks, hyperplasia of the lip, nasolabial fold, melolabial fold, scarring from acne vulgaris, and post-rhinoplasty irregularity. The tissue defect may be treated by introducing a plurality of in vitro cultured autologous fibroblast cells at or proximal to the defect area of the patient's tissue. The autologous fibroblast cells may have been cultured in vitro to expand the number of fibroblast cells in at least one medium that comprises autologous serum. The autologous fibroblast cell cultures may be derived from connective tissue, dermal, fascial fibroblasts, papillary fibroblasts, and / or reticular fibroblasts.

The invention relates to an isolated culture method of piglet myocardial fibroblasts and belongs to the technical field of cell culture in modern biotechnology. The method includes: 1, collecting cardiac tissues from a piglet 1 to 3 days old, shearing ventricular tissues, performing PBS (phosphate buffer saline) pre-cooling, and rinsing several times; 2, digesting the myocardial tissues, namely performing multiple digestion at 37 DEG C with a mixture of trypsin 0.25% and collagenase II 0.1% having a ratio of 1:1, adding DNA deoxyribonuclease (0.02 mg / mL) to reduce cell suspension viscosity and increase cell yield, and adding red blood cell lysis buffer to reduce the amount of red cells; 3, culturing piglet myocardial fibroblasts, namely re-suspending cells with DMEM (Dulbecco modified Eagle medium) high-glucose medium containing fetal calf serum 10-15%, and performing differential attachment to obtain the myocardial fibroblasts. The method is simple and easy to master, time efficient and high in success rate, the obtained piglet myocardial fibroblasts are good in form, good in activity and abundant. Certain basis is provided for establishing a cell model of piglet myocardial fibroblasts to study related diseases such as cardiac hypertrophy in future.

The invention discloses a method for generating and inducing pluripotent stem cells by using pig fibroblasts. The method comprises the following steps: (1) leading cDNA containing pluripotent stem cell factors into the pig fibroblasts; (2) culturing the pig fibroblasts obtained in the step (1); and (3) authenticating pluripotent stem cell clones. The invention successfully induces cells similar to embryonic stem cells (ES) which have the potential of in vitro differentiation and are easy for genetic manipulation. In the invention, the pig induced pluripotent stem (iPS) cells provide convenience for genetic manipulation and can provide scientific data for further developing differentiation research, building disease models, detecting the security of the iPS cells taking the pig as a model, optimizing the inducing strategy of the iPS cells and accelerating the clinic application of human iPS cells.

The invention discloses a method for preparing an autologous triple skin fibroblast preparation. The autologous triple skin fibroblast preparation comprises platelet-rich plasma, autologous collagen concentrate and fibroblasts, wherein the ratio of platelet-rich plasma to the autologous collagen concentrate is 1 to 4; and the concentration of autologous fibroblasts is (1.0-2.0)*10<7> / mL. The preparation method comprises the following steps: separating and culturing human skin fibroblast; preparing the high-concentration autologous collagen solution; preparing autologous platelet-rich plasma; and preparing an autologous triple skin fibroblast injection preparation. Active ingredients of the preparation disclosed by the invention are all from a customer, and are cultured in an autologous serum medium during the cultivation of fibroblasts, so that the preparation is truly safe, effective and individualized. The preparation can be used for skin wrinkle removal, scar repairing, regeneration and other effects, so as to solve a problem of skin aging.

A method for preparing human tissue engineering skin comprises 1, preparation of cutin cell of human skin,2, culturing an preparation of fibroblastic cell, 3, preparation of epidermic cell culture solution and fibroblastic cell culture solution, 4, formation of matrix gel, 5, culturing compound skin, 6, preservation and conveying.

The invention provides a method for constructing a milk cow blood milk barrier three-dimensional model in vitro. The method comprises the steps of cultivation of mammary epithelial cells and fibroblast of a milk cow, cultivation of venous endothelial cells of the milk cow, immunofluorescent identification of the mammary epithelial cells and the venous endothelial cells, cocultivation of Transwell,preparation of a cocultivation medium by taking prolactin, insulin, an epidermal growth factor, cow growth hormone, a vascular endothelial growth factor and a fibroblast growth factor as hormone andcell factors related to growth of the mammary epithelial cells, the fibroblast and the endothelial cells; and finally the epithelial cells, the fibroblast and the endothelial cells are inoculated on atranswell chamber, and accordingly the milk cow blood milk barrier model is successfully constructed. The method can be used for researching a molecular mechanism by which nutrient substances, drugsand toxic substances enter the mammary tissue and the milk in real-time and effectively, not only fills in the gap both domestically and internationally, but also provides a research platform for research on the molecular mechanism by which lipid, vitamins, the drugs and the toxic substances pass through the blood milk barrier.

The invention uses a method for inducing UC-MSC to fibroblasts. According to the method, after in-vitro induction amplification, the fibroblasts are collected to culture supernate, and exosome is obtained by a low temperature ultrafiltration concentration method; the exosome is used as a main active ingredient to prepare a spray. It is proved by experiments that the spray containing fibroblast exocrine can effectively promote the healing of a wound.

The present invention provides a method for production of functional proteins including hormones by renal cells in a three dimensional co-culture process responsive to shear stress using a rotating wall vessel. Natural mixture of renal cells expresses the enzyme 1-a-hydroxylase which can be used to generate the active form of vitamin D: 1,25-diOH vitamin D3. The fibroblast cultures and co-culture of renal cortical cells express the gene for erythropoietin and secrete erythropoietin into the culture supernatant. Other shear stress response genes are also modulated by shear stress, such as toxin receptors megalin and cubulin (gp280). Also provided is a method of treating in-need individual with the functional proteins produced in a three dimensional co-culture process responsive to shear stress using a rotating wall vessel.

The invention provides a method for inducing the transformation of totipotent stem cells into mesenchymal stem cells. In the method, a simple culture method, a fibroblast medium and a monoclonal separation method are adopted to purify and obtain mesenchymal stem cells, the mesenchymal stem cell content is over 95 percent, and more than 109 mesenchymal stem cells can be obtained. The invention has the advantages that: (1) the cells obtained by the method have excellent boundary expansion performance, have properties that can be retained in 15 generation and can from bones, cartilage and muscle tendons; (2) the method for acquiring the cells of the invention is simple, high in efficiency and free from flow type, magnetic bead and the like sorting and purification methods and greatly reduces the cost of the transformation of totipotent stem cells into mesenchymal stem cells; and (3) the method is suitable to be used to repair of defects in tissues such as bone, muscle tendon, cartilage and skin and suitable to be used as seed cells in tissue engineering.

The invention discloses a preparation method and application of autologous fibroblasts. The preparation method of the autologous fibroblasts comprises the following steps that autologous skin tissue of (2.5-3.5 mm)*(2.5-3.5mm) is taken, washed and cut; the cut autologous skin tissue is evenly flatly laid in a culture dish, a fibroblast culture medium is added, and the autologous skin tissue is placed in a culture box with the temperature being 37 DEG C and the CO2 concentration being 5% for primary cell culture; and after the cell confluence degree reaches 70%-80%, trypsin is used for removingepithelial cells in a digestion mode, then digestion is terminated through the fibroblast culture medium, subculture is conducted, and the autologous fibroblasts are obtained. The autologous fibroblasts prepared through the method are high in purity, high in safety and excellent in performance.

A process for inducing fibroblast to become cartilage cell includes culturing the fibroblasts in the culture liquid contaniing the cartilage form generating protein for 3-30 days to generate cartilage cells, and separating them for culture liquid.

The invention discloses a method for culturing chicken embryonic germ cells and a special culture solution thereof. The culture solution is a high sugar DMEM solution containing the following solutes: 3.5 to 4.5 g / L of glucose, 1.5 to 2.5 mM of L-glutamine, (4.5 to 5.5) x 10<5> M of beta-mercaptoethanol, 1 to 1.5 mM of hydroxyethyl piperazine ethyl sulfuric acid, 100 to 120 IU / ml of leukemia inhibitory factor, 4 to 6 ng / ml of stem cell growth factor, 10 to 12 ng / ml of basic fibroblast growth factor, 10 to 12 ng / ml of insulin-like growth factor-1, 10 to 15 percent of fetal bovine serum, 1 to 3 percent of chicken serum, 1 to 1.2 percent of chick embryo extract, and antibiotic. The method for culturing the chicken embryonic germ cells is to culture the chicken embryonic germ cells on a chicken embryonic fibroblast culture layer. Through the reformation and the modification of EG cells, new varieties of chicken can be effectively cultured.

The invention discloses a method and culture media for inducing osteogenic differentiation of an induced pluripotent stem cell of a mouse. The method provided by the invention comprises the steps of inoculating the pluripotent stem cell of the mouse into a differentiation culture medium I to form an embryoid body with enriched mesenchymal cells, then separating the embryoid body and culturing in a differentiation culture medium II to obtain an osteoblast, wherein the differentiation culture medium I disclosed by the invention is formed by adding all-transretinoic acid and basic fibroblast growth factor on the basis of a fibroblast culture medium, and the differentiation culture medium II is formed by reducing the D-glucose content and adding BMP4 or BMP7 on the basis of an osteoblast culture medium. The method and the culture media provided by the invention can significantly promote the osteogenic differentiation of the pluripotent stem cell of the mouse, have the characteristics of short cycle, high differentiation rate, stable effect and the like, and have broad application prospect and practical value.

The invention provides a culture medium. The culture medium comprises a basal medium and a platelet lysate, preferably, comprises gutamic acid and / or proline. The invention also provides a method of culturing the fibroblasts using the above cell culture medium, and an application of the cultured fibroblasts as a raw material for cosmetic or tissue repair.

The invention provides a making method of a three-dimensional vascularized musculocutaneous flap based on biological 3D printing. Coaxial biological 3D printing is utilized; a calcium chloride solution is taken as an inner layer; methacrylic anhydride gelatin, sodium alginate, polyethylene glycol diacrylate and vascular smooth muscle cells are mixed to serve as outer-layer biological ink; a printed scaffold is thoroughly cross-linked with blue light through calcium chloride; a three-in-one perfusate scaffold is finally formed, so that blood supply is established; vascular endothelial cells are poured into the scaffold; vascular cell components are simulated; fibroblasts, adipose tissue-derived stromal cells and skeletal muscle myoblasts are respectively inoculated on a three-layer scaffold; a fibroblast culture medium, an adipogenesis induction culture medium and a myoblast culture medium are respectively added and put into a reactor for culture; and the three-layer scaffold is folded and bond through rat tail collagen to form the stacked scaffold. According to the invention, integrated perfusion of blood flow and independent culture differentiation and assembly of different cell components are realized.

The avian infectious laryngotracheitis virus and fowl pox virus are cultured by using chicken whole embryo fibroblasts. Particularly, the avian infectious laryngotracheitis virus is successfully cultured by using the chicken whole embryo fibroblasts, the avian infectious laryngotracheitis virus and the fowl pox virus cultured by using the chicken whole embryo fibroblasts are obtained, and a combined freeze-drying active vaccine after a proper freeze-drying protectant is added. The combined freeze-drying active vaccine can be inoculated for immunization in a subalar and subcutaneous inoculation manner, so that side effects of the avian infectious laryngotracheitis virus can be alleviated, immune operations can be simplified, stress response can be reduced and production cost can be lowered. At the same time, the freeze-drying protectant prepared by a novel formula can effectively prolong storage time of the active vaccine and provide convenience for storing and transporting the products.

The invention relates to the technical field of biological tissue engineering, and discloses a lung cancer organoid culture solution and a culture reagent combination and a culture method thereof, and the lung cancer organoid culture solution comprises a complete medium and an autogenous condition medium; the complete culture medium comprises a conditioned culture medium, a basic culture medium, compound antibiotics and growth factors; the autogenous condition culture medium is obtained by culturing fiber cells extracted from lung cancer tissues by using a tumor-associated fibroblast culture medium. All the components of the culture solution have a synergistic effect, so that the formation and growth of lung cancer organoid cells are accelerated, and the culture speed is increased while the cell harvesting number is increased; and the in-vitro cultured organoid is more bionic, can be suitable for personalized culture of the lung cancer organoid, and can maintain the physical structure and pathological characteristics of the primary tissue of each patient.

Certain embodiments herein are directed to a method of treating a tissue associated with a defect in a human including wrinkles, rhytids, depressed scar, cutaneous depressions, stretch marks, hyperplasia of the lip, nasolabial fold, melolabial fold, scarring from acne vulgaris, and post-rhinoplasty irregularity. The tissue defect may be treated by introducing a plurality of in vitro cultured autologous fibroblast cells at or proximal to the defect area of the patient's tissue. The autologous fibroblast cells may have been cultured in vitro to expand the number of fibroblast cells in at least one medium that comprises autologous serum. The autologous fibroblast cell cultures may be derived from connective tissue, dermal, fascial fibroblasts, papillary fibroblasts, and / or reticular fibroblasts.