Kit for preparing disease-treating agent, disease-treating agent and method for preparing disease-treating agent
A disease treatment and kit technology, applied in biochemical equipment and methods, chemical instruments and methods, sensory diseases, etc., can solve the problems of not being used to treat heart failure, unable to cope with emergency surgery, cost and time, etc. Not easy to reject, stable and constant effect, easy to commercialize effect
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[0060] (Method for preparing mesenchymal stem cells)
[0061] The mesenchymal stem cells can be prepared by a method known to those skilled in the art. The method for preparing adipose tissue-derived mesenchymal stem cells will be described below as an example. Adipose tissue-derived mesenchymal stem cells can be obtained by, for example, the production method described in US Patent No. 6,777,231, for example, can be produced by a method including the following steps (i) to (iii):
[0062] (i) a process of obtaining a cell suspension by enzymatically digesting adipose tissue;
[0063] (ii) a process of pelleting the cells and resuspending the cells in a suitable medium; and
[0064] (iii) A step of culturing cells on a solid surface and removing cells that do not appear to be bound to the solid surface.
[0065] The adipose tissue used in the step (i) is preferably washed adipose tissue. Washing can be performed by using a physiologically acceptable saline solution (such a...
Embodiment
[0108] The present invention will be specifically described by the following examples, but the present invention is not limited to the examples.
[0109] [1] Preparation of the disease therapeutic agent of the present invention
[0110] Preparation of adipose tissue-derived mesenchymal stem cells
[0111] After obtaining the consent of the human donor, the subcutaneous fat tissue obtained by liposuction was washed with physiological saline. In order to destroy the extracellular matrix and separate the cells, collagenase (Roche) was added (the solvent was physiological saline), and the mixture was dispersed by shaking at 37° C. for 90 minutes. Next, the suspension was centrifuged at 800 g for 5 minutes to obtain a precipitate of interstitial vascular cell population. Serum-free medium for mesenchymal stem cells (Rohto Co.) was added to the above cell pellet, the cell suspension was centrifuged at 400 g for 5 minutes, the supernatant was removed, and then suspended in serum-...
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