The present disclosure discloses a method for preparing a lysate, from a combination of umbilical cord blood (UCB) and maternal blood (MB) derived platelets by obtaining platelet-rich plasma via sedimentation processes and then mixing platelet-rich plasmas so obtained. The present disclosure further discloses a combination of umbilical cord blood and maternal blood platelet lysate (UCB+MB PL), and the PL so obtained shows greater efficacy as cell and tissue culture media supplement over and above its individual components, umbilical cord blood platelet lysate (UCB PL) and maternal blood platelet lysate (MB PL) and especially over foetal bovine serum (FBS). Moreover, the method of the present disclosure provides an economic, environment friendly, ethical and efficacious method to reach a cell and tissue culture supplement with far-reaching therapeutic applications.

Methods, systems, and compositions are provided for extracting bone marrow cells from bone obtained from deceased donors, for preparing the bone marrow for cryopreservation, and for obtaining desired cells from cryopreserved and fresh bone marrow

The present disclosure provides a method of separating hematopoietic stem cells from umbilical cord blood, including the following steps: a hydroxyethyl starch solution is added into cord blood and mixed uniformly, then centrifuged to get an upper liquid layer and a lower erythrocyte layer; the liquid in the upper layer and the erythrocytes in the lower layer are centrifuged respectively; an upper plasma layer and a basal cell layer are obtained after the centrifugation of the upper liquid layer, the cells in the basal layer are resuspended; a superficial buffy-coat layer is obtained after the centrifugation of the erythrocytes, and the superficial buffy-coat layer is metered with the plasma in the upper layer, and then centrifuged to get a lower cell layer, which is precipitated to remove erythrocytes and then resuspended; the above resuspended cells are added into freezing medium, mixed uniformly and then stored in a liquid nitrogen tank.

A three-dimensional culture method for large-scale preparation of stem cells, comprising a three-dimensional microcarrier-based cell resuscitation method, a three-dimensional microcarrier cell culture-based in situ passage method, a three-dimensional microcarrier in situ freeze preservation method for cells, a three-dimensional microcarrier cell adsorption culture method, a method for harvesting cells on a three-dimensional microcarrier, a method for sampling cells cultured on a microcarrier, and a three-dimensional microcarrier-based cell large-scale expansion method.

A production method for pluripotent stem cells is provided, the method including the following (a) and (b): (a) a process filling a culture container with a liquid medium and thereafter raising, the temperature of the liquid medium in the culture container to the temperature at which pluripotent stem cells can proliferate; and (b) a process seeding pluripotent stem cells in the liquid medium in the culture container and culturing the pluripotent stem cells in suspension.

The present invention provides a polynucleotide having the nucleotide sequence as set forth in SEQ ID NO: 1 encoding 9-lipoxygenase. The present invention also provides recombinant plasmid expression vector comprising said polynucleotide. The recombinant protein 9-lipoxygenase encoded by the polynucleotide leads to production of lactones in fruits such as mangoes.

A method for making an acellular human amnion-derived composition configured for therapeutic use is disclosed and generally includes the steps: obtaining amniotic membrane tissue; testing the amniotic membrane tissue for pathogens; washing the amniotic membrane tissue; manually removing blood-containing chorion tissue from the amniotic membrane tissue decellularizing the amniotic membrane tissue with xeno-free enzymes; collecting amniotic cells from the decellularized amniotic membrane tissue; seeding the amniotic cells for culture into xeno-free media formulated for mesenchymal stem cells; growing the amniotic cells to a specified confluency; collecting conditioned media; and freezing the collected conditioned media; wherein the method further includes irradiating the conditioned media.

The specification describes an improved serum-free animal cell culture medium, which can used for the production of a protein of interest. Ornithine, or a combination of ornithine and putrescine can be added to serum-free media or chemically defined media to improve viable cell density, to reduce cell doubling time, and to increase the production of a protein of interest.

Methods for the production of replication-incompetent recombinant vesicular stomatitis virus (rVSV) in suspension cell culture are disclosed. In some embodiments, the methods include inoculating a suspension cell culture medium with packaging cells, transfecting the packaging cells with a plasmid comprising a nucleic acid molecule encoding a viral envelope glycoprotein, introducing rVSV, devoid of a gene encoding a functional envelope glycoprotein, into the suspension cell culture medium, and isolating rVSV produced from the packaging cells with the viral envelope glycoprotein incorporated into its viral envelope.

A method and apparatus of aseptic processing and production of cells in a non-sterile enclosure apparatus without chemical biocides is disclosed, by controlling the level of humidity throughout the enclosure to 25% relative humidity (RH) or less, and preferably 20% or 15% or less RH. In addition, the temperature is controlled to 37° C., and consistent gas flow is maintained the enclosure. Colony forming units from microbial contamination detected by environmental monitoring within the enclosure are significantly reduced in this method.