The present invention relates to, among others, an in vitro method of modifying a cell graft containing immune cells comprising the steps of incubating a cell graft containing immune cells with an anti CD4 antibody wherein said incubating is carried out for from 1 minute to 7 days, b) removing unbound antibody from said graft; as well as to corresponding modified grafts and uses. The invention further relates to the modification of antibodies reactive to the CD4 human leukocyte antigen to provide anti-CD4 antibodies that have a reduced number of potential T-cell epitopes but retain the ability to bind to CD4, such as to an anti human CD4-antibody comprising a heavy chain immunoglobulin variable domain (VH) and a light chain immunoglobulin variable domain (VL), wherein at least one T cell epitope located outside the CDRs of said immunoglobulin variable domains is removed from said immunoglobulin variable domains. Preferably, the specificity and mode of action of the anti-CD4 antibodies are not affected by the modification(s).

A method for isolating and processing bone marrow derived stem cells, including the steps of: (a) collecting a biological sample containing adherent bone marrow stem cells in a receptacle with interior walls coated with a cell-adherent substrate; (b) incubating the bone marrow cells on the adherent substrate so that a layer of adherent bone marrow stem cells adheres to the substrate; (c) washing any non-adherent cells from the substrate; and (d) collecting the bone marrow stem cell layer. Isolation kits and use of bone marrow cells harvested for cell therapies are also described.

The present invention provides a technique which enables organization of bone marrow cells by a simple method in a short period of time.

A method for preparing a bone marrow cell aggregate, comprising adding a liquid containing a bone marrow cell population to a medium containing a swellable material and culturing the bone marrow cell population in the presence of the swellable material. A method for reassembling a bone marrow tissue, comprising adding a liquid containing a bone marrow cell population to a medium containing a swellable material and culturing the bone marrow cell population in the presence of the swellable material.

According to common knowledge in the art, it has been considered difficult to reorganize once disintegrated bone marrow tissue without changing the cell composition (that is, without adding any adherent cell or extracellular matrix which will work as a “connecting material (binder)”). Indeed, it was impossible to aggregate bone marrow cells by conventional methods. As a result of its achievement, the present invention changes such conventional thought and results and provides a major breakthrough technique pertaining to 3D culture of bone marrow cells. It has also been confirmed that culture of a bone marrow-like tissue reassembled by the method of the present could be continued up to day 14 in the MC medium.

A method of producing Klotho protein includes preparing a Klotho plasmid DNA vector, culturing cells, transfecting the cells with the Klotho plasmid DNA vector in a cell culture medium, growing the transfected cells, and harvesting the cell culture supernatant by removing the transfected cells. The Klotho plasmid DNA vector has a mammalian selection marker and a Klotho open reading frame. The cells are primary fibroblast cells and/or mesenchymal stromal cells. A method of manufacturing a cosmetic composition includes combining Klotho protein or the cell culture supernatant with a cosmetically acceptable vehicle. A method of treating a patient to improve the condition and appearance of aging skin includes topically administering the cosmetic composition to the patient. By upregulating the Klotho gene in vitro and incorporating the Klotho protein and growth factors into a composition, transepidermal water loss, skin atrophy, and free radical damage to the skin may be addressed.

Provided herein are methods and systems for bio-printing of three-dimensional organs and organoids. Also provided herein are bio-printed three-dimensional organs and organoids for use in the generation and/or the assessment of immunological products and/or immune responses. Also provided herein are methods and system for bio-printing three-dimensional matrices.

The embodiments disclosed herein generally relate to systems, devices and methods for the fractionation, isolation, extraction and processing of the adipose supernatant layer of a bone marrow aspirate. In particular, the various embodiments relate to systems, devices, and methods of obtaining, utilizing, and processing the adipose supernatant layer of a bone marrow aspirate as a source of mesenchymal stem cells.

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

A method of maintaining cellular viability of harvested bone, where the method includes: providing a source of bone or bone particles; combining the bone or the bone particles with a sterile solution; and storing the bone or the bone particles in the sterile solution until their introduction into a patient.

The invention provides for transgenic donor animals (e.g., pigs) whose cells, tissues and organs have a better long-term survival when transplanted into a human patient. The transgenic donor animal carries one or more human transgenes which is expressed only when the endogenous gene of the donor animal is knocked out shortly before a graft is harvested for transplantation. This “genetic switch” allows the donor animal to remain healthy during the majority of its lifetime, while still allowing expression of the human transgene for optimal transplant tolerance in a human recipient. The transgene may encode a cytokine receptor, an adhesion molecule, or a complement regulatory protein.

Disclosed herein are three-dimensional bone tissue grafts produced from stacked demineralized bone paper. Also disclosed are methods for treating a subject using tissue grafts produced from the disclosed demineralized bone paper. Also disclosed are assay systems that involves culturing bone-promoting cells on the disclosed demineralized bone paper.