A primary culture method in which cells contained in a tissue collected from a living body are primary cultured in vitro, in which the cells in the tissue collected from the living body are seeded and cultured on a top surface of a cell structure containing cells constituting a stroma and composed of a single layer or two or more cell layers laminated in the thickness direction.

The invention relates to an ex vivo generated population of tissue-specific anti-inflammatory macrophages and methods of making and using such macrophages.

Embodiments of the disclosure include methods and compositions for disc repair in a mammal using conditioned media (and/or one or more components therefrom) from fibroblasts that have been de-differentiated and cultured optionally with one or more particular conditions and/or compositions. In specific cases, fibroblasts that have been de-differentiated are exposed to hypoxia, histone deacetylase inhibitor(s), DNA methyltransferase inhibitor(s), or a combination thereof, and the conditioned media therefrom is provided in an effective amount to an individual.

The present invention features assays for co-culturing primary cells while maintaining key biological activities specific to the primary cells. The invention is based, at least in part, on the discovery that compositions and methods for primary cells in a high-throughput co-culture platform, image analysis for distinguishing cells in co-cultures and assays that are suitable for screening of agents in epithelial cells, such as hepatocytes.

In some aspects, this invention provides a method of making a bone marrow-derived tissue-specific stem cell proliferation, expansion, isolation and rejuvenation extracellular matrix. In other aspects, this invention provides a method of making a tissue-specific fibroblast-derived stem cell differentiation extracellular matrix. Also provided are methods of using such a cell-derived preservation or differentiation matrices to induce tissue-specific differentiation of pluripotent cells, repair damaged tissue, and treat a subject having a physiologic deficiency using the same.

The present disclosure provides a non-autologous product that is a mixture of two or more cells or tissue cultures of fibroblast, or extracts from cultures, or media cultures, isolated from separate individuals, either homogeneous or heterogeneous. The cells or factors are blended together in a product that imparts desired characteristics to the skin of a recipient who is not a source of the mixture. The present disclosure also relates to methods of making and using and/or culturing the fibroblasts including to optimize the potency of the mixture to impart one or more the desired characteristics to the skin of a recipient.

A cell sheet for gene delivery is disclosed. Unlike conventional cell sheets for tissue regeneration, the disclosed cell sheet can be used as a local gene delivery system. Particularly when a virus is used as a gene delivery system, the virus can be proliferated within the cell sheet and acts topically within a therapeutic region. Thus, the cell sheet is superior in the prevention or treatment of cancer, the prevention of cancer recurrence or cancer metastasis, particularly the treatment of multifocal tumor even though the virus dose is remarkably lowered compared to the systemic administration or intratumoral injection of the virus.

An in vitro microfluidic organ-on-chip device is described herein that mimics the structure and at least one function of specific areas of the epithelial system in vivo. In particular, a stem cell-based Lung-on-Chip is described. This in vitro microfluidic system can be used for modeling differentiation of cells on-chip into lung cells, e.g., a lung (Lung-On-Chip), bronchial (Airway-On-Chip; small-Airway-On-Chip), alveolar sac (Alveolar-On-Chip), etc., for use in modeling disease states of derived tissue, i.e. as healthy, pre-disease and diseased tissues. Additionally, stem cells under differentiation protocols for deriving (producing) differentiated lung cells off-chips may be seeded onto microfluidic devices at any desired point during the in vitro differentiation pathway for further differentiation on-chip or placed on-chip before, during or after terminal differentiation. Additionally, these microfluidic stem cell-based Lung-on-Chip allow identification of cells and cellular derived factors driving disease states in addition to drug testing for diseases, infections and for reducing inflammation effecting lung alveolar and/or epithelial regions. Further, fluidic devices are provided seeded with primary alveolar cells for use in providing a functional Type II and Type I cell layer, wherein Type II cells express and secrete surfactants, such as Surfactant B (Surf B; SP-B) and Surfactant C (Surf C; SP-C), which were detectable at the protein level by antibody staining in Type II cells. A number of uses are contemplated for the devices and cells, including but not limited to, for use under inflammatory conditions, in drug development and testing, and for individualized (personalized) medicine. Moreover, an ALI-M was developed for supporting multiple cell types in co-cultures with functional Type II and Type I cells.

The present invention relates generally to compositions for wound closure. More specifically, the present invention provides human skin equivalents engineered to express exogenous polypeptides (e.g., antimicrobial polypeptides and keratinocyte growth factor 2) and compositions and methods for making human skin equivalents engineered to express exogenous polypeptides. In addition, the present invention provides methods for treatment of wounds with human skin equivalents engineered to express exogenous polypeptides.

Described herein are methods for providing an in vitro intestinal model system, e.g., using primary cells instead of cell lines and/or cancerous cells.