Method for detecting length of the shortest telomere in cells by using improved STELA method
A cell and short-terminal technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., to achieve the effect of enhancing the proliferation ability
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0064] Example 1. Cell culture
[0065] Cell culture: Human skin fibroblasts BJ and human lung fibroblasts IMR-90 (ATCC) were cultured in EMEM medium supplemented with 10% BSA. The BJ cell line was cultured continuously until it became senescent, during which time cells and gDNA were collected weekly. The IMR-90 cell lines were collected at four different passage numbers (PD 19.5, PD 27, PD 36, PD 44) and cryopreserved, and the remaining cells were continuously cultured until senescence.
Embodiment 2
[0066] Example 2. β-Gal cell staining
[0067] BJ cells were seeded on glass slides, washed twice with 1x PBS; fixed with 3.7% formaldehyde at room temperature for 5 min; after washing with 1x PBS, freshly prepared β-gal solution (X-Gal 1 mg / ml, citric acid 40 mM, pH 6.0, potassium ferrocyanide 5 mM, potassium ferricyanide 5 mM, sodium chloride 150 mM, magnesium chloride 2 mM) stained cells, and cultured overnight at 37 °C. Observe staining of at least 200 cells under a microscope.
Embodiment 3
[0068] Example 3. Separation of growing and senescent cells by CiI cell staining and flow cytometry
[0069] After BJ cells and IMR-90 cells were trypsinized, they were suspended and stained in 5 uM DiI solution (purchased from Molecular Probes) for 20 min. After staining, wash twice with 1x PBS, and inoculate in culture dishes at a density of about 2000 cells / cm 2 . The cells were cultured in the dark for 5-7 days. After trypsinization, the cells were washed twice with freshly prepared PBS buffer containing 2% PBS and 0.02% EDTA. Then the cells were collected and suspended in the same buffer, and placed in on ice.
[0070] Cells in growth phase and senescent cells were separated by flow cytometry using BD FACSAria I Cell-Sorting System (BD Bioscience). Growth phase cells were stained by DiI to a low degree, and senescent cells were vice versa. Using the DiI signal as a reference, the long-term cells (PROL) and senescent cells (SEN) were separated and collected in test tub...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More - R&D
- Intellectual Property
- Life Sciences
- Materials
- Tech Scout
- Unparalleled Data Quality
- Higher Quality Content
- 60% Fewer Hallucinations
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com