30 results about "Cryopreserved Tissue" patented technology

A bioreactor and methods of using same for making tissue constructs and for conditioning tissue-engineered constructs and harvested tissues such as cryopreserved tissues. The bioreactor allows for static and dynamic culture / conditioning. The bioreactor is dual chambered (one chamber above and one below the cells or construct) to allow for application of biochemical and / or biomechanical stimuli to each side of the cells / construct.

The present invention relates to a method and composition for the cryopreservation of adipose tissue with the intention to use this tissue in the culturing of stem and / or progenitor cells. The method uses a specific cryoprotection medium to prevent damage of the original tissue during the cryopreservation while still maintaining a high viability of the stem and / or progenitor cells obtained from the cryopreserved adipose tissue. Furthermore the cryoprotection medium of the present invention does not contain any kind of xenogeneic sera, a critical factor since it is the intention of that the cryopreserved tissue is used for obtaining stem and / or progenitor cells that can be used in medicine. The cryoprotection medium is characterized in that it is a solution of physiological water comprising glycerol and sucrose and / or trehalose and optionally serum albumin.

A bioreactor and methods of using same for making tissue constructs and for conditioning tissue-engineered constructs and harvested tissues such as cryopreserved tissues. The bioreactor allows for static and dynamic culture / conditioning. The bioreactor is dual chambered (one chamber above and one below the cells or construct) to allow for application of biochemical and / or biomechanical stimuli to each side of the cells / construct.

The invention belongs to the technical field of biology, and discloses a method for preparing mesenchymal stem cells. The method takes a neonatal umbilical cord as a raw material and comprises the following steps: pretreating the umbilical cord; cryopreserving tissues; resuscitating the tissues; separating primary mesenchymal stem cells; and amplifying the mesenchymal stem cells. The method disclosed by the invention has the advantages being easy for industrialization and operation and the like, and can be used for cryopreserving a large quantity of umbilical cord tissues and resuscitating the umbilical cord tissues for clinical need to prepare the mesenchymal stem cells. Compared with an existing method for directly separating the mesenchymal stem cells and cryopreserving the mesenchymal stem cells, the method disclosed by the invention can be used for sufficiently preserving the most original mesenchymal stem cells contained in the umbilical cord tissues, thereby greatly reducing the preservation cost and preventing the mesenchymal stem cells from influencing clinical application after being differentiated due to multiple-time passage in vitro.

The application provides a tissue and / or cell cryopreservation protective solution as well as preparation and application thereof. The tissue and / or cell cryopreservation protective solution comprisesa permeating protective agent and a non-permeating protective agent, wherein the cryopreservation protective solution further comprises platelet lysates. In addition, the application further providesa method for cryopreserving tissues and / or cells by using the tissue and / or cell cryopreservation protective solution provided by the application, as well as a cryopreserved tissue and / or cell and use of the cryopreserved tissue and / or cell.

The invention discloses a cryopreservation method of umbilical cord tissue blocks, and relates to a cryopreservation method of umbilical cords. The invention aims at solving the problems that the preservation time of umbilical cord tissues is short, and the cell death of the umbilical cord tissues is easily caused at present. The method comprises the following steps: 1, carrying out treatment of the tissue blocks, namely in a sterile operating platform, carrying out disinfection treatment on an umbilical cord, cleaning the umbilical cord normal saline, shearing the umbilical cord into segments, peeling off a wharton jelly, placing the wharton jelly in a clean centrifuging tube and cutting the wharton jelly into pieces; and 2, adding DMEM in the tissue blocks, resuspending the tissue blocks, then adding a pre-cooling cryopreservation liquid, sub-packaging the tissue blocks in cryopreservation tubes, cooling by use of a program-controlled cooler, and transferring the tissue blocks in liquid nitrogen for long-time storage after bacterial detection, microbiological detection and virus detection are all negative, so as to finish the cryopreservation of the umbilical cord tissue blocks. The directly-inoculated adherent survival rate of the cryopreserved and unfrozen tissue blocks in culture bottles is more than 98%, the cell growth is good, and moreover, an osteogenic differentiation experiment carried out on the cultured cells of the cryopreserved tissue blocks indicates that the differentiation function is high. The method is used for storing the umbilical cord tissues.

A method for cryopreservation of a tissue equivalent whereby the viability of frozen cells and the biological activity of thawed cells are improved and the steps are simplified; and a cryopreserved tissue equivalent obtained by the method. Cells suspended in a cryopreserving solution are inoculated into a matrix and then frozen before the cells adhere to the matrix.

The invention relates to a solution system for promoting cryopreserved tissue organ and cell activity recovery. One or several kinds of materials from water and polyalcohol are used as basic solutions; polyphenol, citric acid, poloxamer and vitamin E are added into the basic solutions; tissue organs and cells are put into the solution system for cryopreserving; after the cryopreserving, ultrasonicwaves with the wavelength in the range of 20KHz to 1MHz and the sound intensity not greater than 3W / cm<2> are used for performing irradiation treatment on cells; by regulating the proportion of different ingredients in the solution and controlling the wavelength, sound intensity and irradiation treatment of the ultrasonic waves, the biological activity after the cryopreserved tissue organ and cell recovery is improved.

An method for enhanced follicular tissue transplantation and management is provided. Follicular tissue is first obtained from a donor during a dedicated harvesting procedure or as a by-product of another cosmetic surgery procedure, then cryopreserved and stored. When implantation is desired, the cryopreserved tissue is thawed, and the tissue that survived the cryopreservation and thawing processes is implanted in the recipient. In another aspect of the invention, the harvesting process is conducted when the quality of donor follicular tissue is as optimal as possible, and is stored in cryopreserved form until the donor requires implantation thereof, such that the newly implanted thawed follicular tissue is of higher quality than the recipient's current follicular tissue. In yet another aspect of the present invention, a method for managing the business aspects of the transplantation process, including harvested follicular tissue grading, cataloguing and banking is provided.

The invention provides Sepia pharaonis neuropeptide GnRH and a use thereof. A preparation method of the Sepia pharaonis neuropeptide GnRH comprises 1) pretreatment: taking a living Sepia pharaonis, dissecting to take the brain tissue and storing the brain tissue in a preservation liquid at an ultralow temperature of -70 to -90 DEG C, 2) supernatant collection: taking the brain tissue, adding the equal volume of normal saline into the brain tissue, carrying out homogenization and centrifugation, taking the supernatant A, then adding the equal volume of distilled water into the supernatant A, carrying out centrifugation, taking the supernatant B, and merging the supernatants A and B to obtain a supernatant C, and 3) column chromatography: carrying out column chromatography on the supernatant C, carrying out color development, collecting components and carrying out freeze drying. The preparation method can realize cryopreservation of the tissue, realizes excellent effects, and can effectively prevent the degradation of the Sepia pharaonis brain tissue. The extract can be used in drugs for prevention or treatment on hormone-dependent diseases.

The invention discloses a preparation and cryopreservation method of human placental chorionic tissues. The method comprises the following steps: (1) separation and cryopreservation of a placental amnion and a decidua; (2) separation and cryopreservation of placenta subchorionic large vascular tissues; 3) separation and cryopreservation of the placental chorionic tissues; and (4) treatment and cryopreservation of the placental chorionic tissues. The invention also provides a method for resuscitation after cryopreservation. The cryopreservation method facilitates the improvement of the activityof cryopreserved tissues and cells, the morphology, the function and the structure of resuscitated cryopreserved tissues are same to those of fresh tissues, the overall survival rate of cells in thetissues reaches 90% or above, and the preserved tissues can be used in the field of separation of stem cells and epithelial cells, and also can be used in the field of tissue engineering transplantation.

The invention provides a deep cryogenic freezing method for tumor tissue blocks, and relates to tumor tissue freezing methods. The deep cryogenic freezing method is used for solving the problems of the existing tissue cryopreservation methods that the activity of resuscitated tumor cells is remarkably lowered and the tumor forming rate is low. The method comprises the following steps: (1) preparing a cryopreservation solution; (2) treating and freezing the tissue blocks; (3) resuscitating cryopreserved tumor tissue blocks. According to the method, tumor tissue cryopreservation is carried out by using a permeable and impermeable cryopreservation protectant combined method, meanwhile, a preparing proportion of reagents is optimized, the proportion of DMSO in the cryopreservation solution is lowered to 2%, the cytotoxicity of tissue is lowered, a very good protective action on the cryopreserved tissue is achieved, thus, the cryopreserved tissue has relatively high mouse tumor forming performance after resuscitation, and the tumor forming rate reaches 90% or more. The method is applied to the freezing of the tumor tissue.

A cryopreservation medium and a cryopreservation method which make it possible to cryopreserve tissues and cells with high viability are provided. A cryopreservation medium contains polyphenol at a concentration of from 30 to 120 ppm. In an embodiment, a cryopreservation method for tissues and cells comprises: incubating tissues or cells for 2 hours in culture medium supplemented with polyphenol at concentrations of from 30 to 120 ppm; suspending tissues or cells in a cryopreservation medium; freezing the suspension of the tissues or cells in the cryopreservation medium; and storing the frozen suspension of tissues or cells in the cryopreservation medium.

The invention discloses a systematic method for preparing and cryopreserving placental tissues according to a structure layer. The method comprises the following steps: (1) separation and cryopreservation of a placental amnion and a decidua; (2) separation and cryopreservation of placenta subchorionic large vascular tissues; 3) separation and cryopreservation of the placental chorionic tissues; and (4) treatment and cryopreservation of the placental chorionic tissues. The invention also provides a method for resuscitation after cryopreservation. The placental amnion, the decidua, the subchorionic large vascellum, the chorion and other tissues are respectively separated and preserved, so the improvement of the activity of cryopreserved tissues and cells is facilitated, the morphology, the function and the structure of resuscitated cryopreserved tissues are same to those of fresh tissues, the overall survival rate of cells in the tissues reaches 90% or above, and the preserved tissues can be used in the field of separation of stem cells, and also can be used in the field of tissue engineering transplantation. The systematic method for preserving various tissues of the complete placenta is provided to provide a great biological source for the studying of placenta-derived stem cells.

The invention discloses a preparation cryopreservation method of the human placental subchorionic villi aorta tissue. The method comprises the steps that (1) a placenta is washed cleanly; the abscisicdecidua is cut off along the edge of the placenta, and the amniotic membrane is removed; (2) the placenta fetal surface chorion with the amniotic membrane and the aortas are separated and washed withnormal saline or PBS buffer liquid, the blood vessels are washed so as to remove the stranded blood clots in the blood vessels; and then, the aortas connected with the chorion lamina membranacea is cut off one by one; and (3) the blood vessels are transferred into a cryopreserved pipe or a cryopreserved bag, vitrified cryopreservation liquid is guided through a three-step method, the blood vessels are transferred into a programmed freezer so as to be cooled at -80 - -90 DEG C, and then the blood vessels are transferred to liquid nitrogen cryopreservation. The cryopreservation method is beneficial for improvement of the activity of the cryopreserved tissue and the cells, the morphology, the function and the structure are consistent with the fresh tissue after being recovered, the preservedtissue can not only be applied to fields of separation of stem cells and epithelial cells, but also can be applied to fields of tissue engineering transplantation.

The application provides a tissue and / or cell cryopreservation protective solution as well as preparation and application thereof. The tissue and / or cell cryopreservation protective solution comprisesa permeating protective agent and a non-permeating protective agent, wherein the cryopreservation protective solution further comprises platelet lysates. In addition, the application further providesa method for cryopreserving tissues and / or cells by using the tissue and / or cell cryopreservation protective solution provided by the application, as well as a cryopreserved tissue and / or cell and use of the cryopreserved tissue and / or cell.

The invention discloses a protective solution as well as a preparation method and application thereof. The protective solution comprises a cryoprotectant and a complete culture medium, the cryoprotectant is composed of a permeable protective agent and a non-permeable protective agent, and the non-permeable protective agent contains beta-1, 3 glucan; the complete culture medium consists of a serum-free basal culture medium, a serum substitute and L-glutamine; and the protective solution does not contain fetal bovine serum, autologous cord blood serum and human serum albumin. The protective solution can provide a good cryopreservation environment for tissue cells, beta-1, 3 glucan can improve the stability of the protective solution and improve the cryopreserved tissue protection effect andthe cell viability after resuscitation, the complete culture medium provides nutrition for umbilical cord tissues and cells and is consistent with a culture medium for stem cell resuscitation and culture, so that the convenience in preparation and application of the protective solution is improved. The protective solution has the advantages of being convenient to use, short in preparation period of a cryopreservation preparation, low in cost, safe and reliable in clinical application and the like.

The invention discloses a preparation and cryopreservation method of human placental chorionic tissues. The method comprises the following steps: (1) cleaning a placenta, and cutting off a shed decidua along the edge of the placenta to remove the amnion; and (2) separating the amnion-removed placenta fetus chorion and large vessels to obtain remaining placenta tissues which are placental chorionictissues, cutting the placental chorionic tissues to form small blocks, cutting the small blocks to form slices, placing the slices in a cryopreservation bag or a cryopreservation tube, guiding a vitrification cryopreservation solution through a three-step process, transferring the obtained mixture into a programmable cooler, reducing the pressure to -80 to -90 DEG C, transferring the cooled mixture to liquid nitrogen, and performing cryopreservation. The cryopreservation method facilitates the improvement of the activity of cryopreserved tissues and cells, the morphology, the function and thestructure of resuscitated cryopreserved tissues are same to those of fresh tissues, the overall survival rate of cells in the tissues reaches 90% or above, and the preserved tissues can be used in the field of separation of stem cells and epithelial cells, and also can be used in the field of tissue engineering transplantation.

This invention is directed to an efficient cryopreservation package design of harvested mammalian tissues and living cultured tissue equivalents made by in vitro technology. The invention involves immersing a mammalian tissue or cultured tissue equivalent in a cryoprotectant solution, agitating the cryoprotectant solution and the immersed tissue to achieve effective penetration of the cryoprotectant solution into the tissue, and then freezing the tissue at a very slow freezing rate. In the freezing step, extracellular ice formation is initiated by seeding. The cryopreserved tissue may be stored for indefinite periods of time prior to use. The cultured tissue equivalent is an in vitro model of the equivalent human tissue, such as skin or cornea, which, when retrieved from storage can be used for transplantation or implantation in vivo or for screening compounds in vitro.

A solution system for promoting recovery of cryopreserved tissues and organs and cell activity of the present invention uses one or more of water and polyols as the base solution, and polyphenols, citric acid, and poloxamer are added to the base solution and vitamin E, put tissues, organs and cells in this solution system for cryopreservation, and apply the wavelength within the range of 20KHz-1MHz and the sound intensity is not greater than 3W / cm after cryopreservation 2 Cells are irradiated by ultrasonic waves. By adjusting the ratio of different components in the solution, and controlling the wavelength, sound intensity and irradiation time of ultrasonic waves, the biological activity of frozen tissues, organs and cells after recovery can be improved.

The invention discloses a method for preparing and freezing human placental subchorionic macrovascular tissue, comprising: (1) washing the placenta; cutting off the decidua along the edge of the placenta to remove the amniotic membrane; (2) removing the amniotic membrane from the placenta The fetal facial chorion and large blood vessels are separated together, washed with normal saline or PBS buffer, and the blood vessels are flushed to remove blood clots in the blood vessels; then, the large blood vessels connected to the chorionic plate are cut off one by one; (3) the blood vessels are transferred Put it into a cryopreservation tube or a cryopreservation bag, introduce the vitrification solution in a three-step method, transfer it to a programmed cooling device, cool it down to -80°C to -90°C, and transfer it to liquid nitrogen for cryopreservation. The cryopreservation method of the present invention is beneficial to improve the activity of cryopreserved tissues and cells, and the morphology, function, and structure after recovery are consistent with those of fresh tissues. The preserved tissues can not only be used in the fields of separating stem cells and epithelial cells, but also in tissue Engineering transplantation and other fields.

The invention discloses a long-term cryopreservation and recovery method for animal tissues, clinical tissues and biopsy samples, and belongs to the technical field of animal cell cryopreservation. The cryopreservation method comprises the following steps: cleaning fresh tissues, and cryopreserving with a special cryopreservation solution; the resuscitation method comprises the following steps: thawing cryopreserved tissues in a water bath, rinsing with a special resuscitation solution, treating with a red blood cell lysis solution, centrifuging, rinsing with a resuscitation solution, and finally obtaining resuscitated tissue cells. According to the method provided by the invention, very good cryopreservation and resuscitation effects can be realized aiming at different types of animal tissue cells; after being cryopreserved for at most two years, the strain still has very good passage activity after resuscitation.

The invention discloses a systematic method for preparing and freezing placental tissue according to structural levels, including: (1) separation and freezing of placental amniotic membrane and decidua; (2) separation and freezing of placental subchorionic large blood vessel tissue ; (3) Isolation and cryopreservation of placental chorionic tissue; (4) Processing and cryopreservation of placental chorionic tissue. The invention also provides a recovery method after cryopreservation. The present invention separates and preserves tissues such as placental amniotic membrane, decidua, subchorionic blood vessels, and chorion, which is beneficial to improving the activity of frozen tissues and cells. The overall survival rate of inner cells is over 90%, and the preserved tissues can not only be used in fields such as separating stem cells, but also in fields such as tissue engineering and transplantation. The invention provides a systematic method for preserving various tissues of the complete placenta, and provides important biological resources for the research on stem cells derived from the placenta.

The invention discloses a cryopreservation method of umbilical cord tissue blocks, and relates to a cryopreservation method of umbilical cords. The invention aims at solving the problems that the preservation time of umbilical cord tissues is short, and the cell death of the umbilical cord tissues is easily caused at present. The method comprises the following steps: 1, carrying out treatment of the tissue blocks, namely in a sterile operating platform, carrying out disinfection treatment on an umbilical cord, cleaning the umbilical cord normal saline, shearing the umbilical cord into segments, peeling off a wharton jelly, placing the wharton jelly in a clean centrifuging tube and cutting the wharton jelly into pieces; and 2, adding DMEM in the tissue blocks, resuspending the tissue blocks, then adding a pre-cooling cryopreservation liquid, sub-packaging the tissue blocks in cryopreservation tubes, cooling by use of a program-controlled cooler, and transferring the tissue blocks in liquid nitrogen for long-time storage after bacterial detection, microbiological detection and virus detection are all negative, so as to finish the cryopreservation of the umbilical cord tissue blocks. The directly-inoculated adherent survival rate of the cryopreserved and unfrozen tissue blocks in culture bottles is more than 98%, the cell growth is good, and moreover, an osteogenic differentiation experiment carried out on the cultured cells of the cryopreserved tissue blocks indicates that the differentiation function is high. The method is used for storing the umbilical cord tissues.