Pulmonary fibrosis treatment agent, ptprr expression-promoting agent, and kit for pulmonary fibrosis treatment
An expression enhancer, pulmonary fibrosis technology, applied in the field of pulmonary fibrosis therapeutic agents, can solve the problems of IPF patients not receiving treatment effect, insufficient donors, inability and so on
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
preparation example Construction
[0046] (Method for preparing mesenchymal stem cells)
[0047] Mesenchymal stem cells can be prepared by methods 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):
[0048] (i) a process of obtaining a cell suspension by enzymatically digesting adipose tissue;
[0049] (ii) a process of pelleting the cells and resuspending the cells in a suitable medium; and
[0050] (iii) A step of culturing cells on a solid surface and removing cells that do not appear to be bound to the solid surface.
[0051] 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 as pho...
Embodiment 1
[0100] Serum-free medium for mesenchymal stem cells (Rohto) was added to commercially available cells (PT-5006 manufactured by Lonza), and 400 g of the cell suspension was centrifuged for 5 minutes, and the supernatant was removed, and then suspended in mesenchymal stem cells. Cells were inoculated into flasks using serum-free medium (Rohto Company) for mesenchymal stem cells. Incubate the cells at 37°C, 5% CO 2 subculture for several days. Several days later, the culture was washed with PBS to remove blood cells and adipose tissue residues contained in the culture medium, and mesenchymal stem cells adhered to the plastic container were obtained.
[0101] The obtained adipose tissue-derived mesenchymal stem cells were dispensed into centrifuge tubes, and centrifuged at 800 g for 5 minutes to obtain cell pellets. After removing the supernatant, an appropriate amount of cell cryopreservation solution (STEM-CELLBANKER (Zenoaq)) was added for suspension. This cell suspension so...
Embodiment 2
[0103]Specimens were collected from cases in which pulmonary fibrotic lesions were indicated and underwent surgery due to lung cancer. The parts where no fibrosis was observed (fibrosis (-)) and the parts where fibrosis was observed (fibrosis (+)) were separated and the tissues were minced separately, and the DMEM medium (manufactured by Wako Co., Ltd.) supplemented with 15% serum was used to separate the ) at 37°C, 10% CO 2 Culture was started in 1, 1, 2, 4, and 7 days, and the medium was replaced after washing with PBS (manufactured by Wako Co., Ltd.), and cultured for a total of 10 days to obtain UIP (Usual interstitial pneumonia; Usual Interstitial Pneumonia) patients. Lung fibroblasts (hereinafter referred to as "UIP cells").
[0104] UIP cells (5.0 × 10 4 cells / well) were seeded into each well of a 24-well plate in CO 2 Incubator (37°C, 10% CO 2 ) overnight, and the culture medium was replaced with DMEM medium supplemented with 1% serum. The above subcultured and cr...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More