Otsuka Pharmaceutical Factory, Inc.

Jichi Medical University

Tamaki Wada

Masako Doi

Masuhiro Nishimura

Eiji Kobayashi

Risa Okairi

Yasutaka Fujita

Norihiro Yoshinaga

Yasuhiro Fujimoto

Takeshi Kikuchi

Takumi Teratani

Chikage Shirakawa

Natsuki Watanabe

A liquid for cryopreserving a cell and a liquid for administration of a mammalian cell capable of cryopreserving a mammalian cell and effectively suppressing cell death after thawing, and a method for cryopreserving a mammalian cell using the cell cryopreservation liquid. The liquid is an isotonic solution that includes 2.0 to 6.0% (w/v) of trehalose or a derivative thereof, or a salt of the trehalose or the derivative, 4.0 to 7.0% (w/v) of dextran or a derivative thereof, or a salt of the dextran or the derivative, and DMSO or glycerin.

Mammalian cell cryopreservation liquid

An object of the present invention is to provide a method capable of inexpensively and conveniently preparing cells having pluripotency and a very low risk of tumorigenic transformation. The cells can be prepared by suspension-culturing mammalian mesenchymal stem cells such as human mesenchymal stem cells from bone marrow (hMSC-BM) and human adipose tissue-derived mesenchymal stem cells (hAT-MSC) (also referred to as 'human adipose-derived stem cells [hADSC]'), 7 types of human adherent mature cells (human hepatocyte cells [hHEP cells], human umbilical vein endothelial cells [HUVEC cells], human dermal lymphatic microvascular endothelial cells [HMVEC cells], human epidermal keratinocyte cells [NHEK cells], human bronchial epithelial cells [NHBE cells], human melanocyte cells [NHEM cells], and human smooth muscle cells [UASMC cells]), and 3 types of human adherent precursor cells (human dermal fibroblast cells [NHDF cells], human skeletal muscle myoblast cells [HSMM cells], and human osteoblast cells [NHOst cells]) to form cell masses (spheroids).

Method for preparing pluripotent stem cells

The problem of the present invention is to provide a method for preserving mammalian cells over a long period of time using a solution for cell transplantation, capable of effectively suppressing cell death when the mammalian cells have been preserved, and the solution for cell transplantation. The present invention is characterized in that mammalian cells are preserved in a physiological aqueous solution for cell transplantation, comprising 2.0 to 6.0% (w/v) of trehalose, a derivative thereof, or a salt of trehalose or the derivative (a trehalose) and 4.0 to 7.0% (w/v) of dextran, a derivative thereof, or a salt of dextran or the derivative (a dextran). The effects of a trehalose and a dextran contained in the physiological aqueous solution for cell transplantation can suppress a decrease in the cell survival rate when mammalian cells are preserved for a long period of time (at least 14 days).

Trehalose and dextran-containing solution for transplanting mammalian cells

A mammalian cell suspension prevents pulmonary embolism formation during administration of mammalian cells, such as mammalian stem cells, through a blood vessel, and a preventive agent against pulmonary embolism formation during administration of mammalian cells through a blood vessel. suspending mammalian cells, such as mammalian stem cells, are suspended in a physiological aqueous solution containing trehalose or its derivative, or a salt thereof as an active ingredient to prepare a mammalian cell suspension for preventing pulmonary embolism formation during administration of the mammalian cells through a blood vessel, including the mammalian cells and trehalose or its derivative, or a salt thereof as active ingredients. Examples of the mammalian cells can include pancreatic islet cells, dendritic cells, natural killer cells, alpha/beta T cells, gamma/delta T cells, and cytotoxic T lymphocytes in addition to mammalian stem cells. Preferred mammalian stem cells can include mammalian mesenchymal stem cells and mammalian pluripotent stem cells.

Trehalose-containing mammalian cell suspension for prevention of pulmonary embolism formation

The present invention provides a mammalian stem cell suspension containing mammalian stem cells and at least one polysaccharide such as trehalose, and the like; a mammalian stem cell aggregation inhibitor containing polysaccharide such as trehalose, and the like; a method of suppressing aggregation of mammalian stem cells, containing suspending the mammalian stem cells in an aqueous physiological solution containing polysaccharide; an inhibitor of a decrease in the survival rate of mammalian stem cells containing polysaccharide such as trehalose and the like; a method of suppressing a decrease in the survival rate of mammalian stem cells, containing suspending the mammalian stem cells in an aqueous physiological solution containing polysaccharides, and the like.

Stem cell suspension

An object of the present invention is to provide a method capable of inexpensively and conveniently preparing cells having pluripotency and a very low risk of tumorigenic transformation. The cells having pluripotency and a very low risk of tumorigenic transformation can be prepared by suspension-culturing mammalian mesenchymal stem cells such as human mesenchymal stem cells from bone marrow (hMSC-BM) and human adipose tissue-derived mesenchymal stem cells (hAT-MSC) (also referred to as 'human adipose-derived stem cells [hADSC]'), 7 types of human adherent mature cells (human hepatocyte cells [hHEP cells], human umbilical vein endothelial cells [HUVEC cells], human dermal lymphatic microvascular endothelial cells [HMVEC cells], human epidermal keratinocyte cells [NHEK cells], human bronchial epithelial cells [NHBE cells], human melanocyte cells [NHEM cells], and human smooth muscle cells [UASMC cells]), and 3 types of human adherent precursor cells (human dermal fibroblast cells [NHDF cells], human skeletal muscle myoblast cells [HSMM cells], and human osteoblast cells [NHOst cells]) to form cell masses (spheroids).

Methods of washing adherent cells, capable of effectively suppressing cell death due to proteolytic enzyme treatment for detaching the adherent cell from a culture vessel and subsequent cell treatment; cell-washing solutions used for the washing method; methods of producing cell suspensions for transplantation using the cell-washing solution; and kits comprising the cell-washing solution. Trehalose or its derivative or a salt thereof is added to physiological aqueous solutions to prepare cell-washing solutions containing trehalose or its derivative or a salt thereof as an active ingredient. The cell-washing solutions can be used to wash adherent cells before detaching the adherent cells from a culture vessel by proteolytic enzyme treatment to suppress cell death due to the proteolytic enzyme treatment. The concentration of trehalose applied to the cell-washing solution may be a concentration capable of suppressing the cell death due to the proteolytic enzyme treatment, such as 0.1 to 20 (w/v) %.

Growing community of inventors

Tokushima, Japan

Tamaki Wada