Angiogenesis is closely associated with wound healing, inflammation, rheumatoid arthritis, diabetic retinopathy, macular degeneration, and tumor growth. During the growth of an organism, angiogenesis is an important part of new tissue growth. In adulthood, except for periodic events that are strictly regulated in special organs, almost every normal tissue lacks a large amount of physiological angiogenesis due to the balance between pro-angiogenic and anti-angiogenic endogenous factors. When the balance is disrupted in the direction of promoting angiogenesis, microvascular endothelial cells (ECs) will shift to an angiogenic phenotype, thereby initiating an angiogenic response, which can be eliminated or accelerated.
In the study, the traditional in vivo angiogenesis experimental methods include corneal micropocket, mesentery, sponge/matrix implants, disc analysis (DAS), zebrafish, etc. However, these experiments are not only technically demanding, costly, and time-consuming, but also involve ethical issues. Compared with in vivo angiogenesis experiments, it is more economical and practical to use a special matrix to maintain cell angiogenesis in vitro. The most commonly used cell growth - maintaining matrix is a soluble basement membrane preparation extracted from EHS mouse tumors - matrix gel.
The main components of matrix gel are laminin, type IV collagen, heparan sulfate proteoglycan (HSPG), and nestin, and it also contains growth factors such as TGF-beta, EGF, IGF, FGF, tissue plasminogen activator, and other growth factors contained in EHS tumors. It provides support, tensile strength, and scaffold support for tissues and cells, and can also serve as a three - dimensional substructure for cell adhesion and movement, as well as a reservoir for growth factors, chemokines, and cytokines. It can also act as a signal for cell morphogenesis and differentiation. Yeasen has developed and produced CeturegelTM matrix gel, which is free of LDEV (lactate dehydrogenase - elevating virus), has ultra - low endotoxin content, and has been tested for mycoplasma to ensure no mycoplasma contamination. It includes different types of matrix gel such as basic concentration, high concentration, and low growth factor.
Product Features
High safety: LDEV (Lactate Dehydrogenase Enhancement Virus) free
Concentration Versatility: Concentration range of 8~20 mg/mL
Good batch stability: Strict production and quality control process to ensure stable performance between batches
Low endotoxin: endotoxin content ≤4 EU/mL
Contaminant detection: no mycoplasma, bacteria and fungi residues detected.
Angiogenesis experiment
- Matrix gel preparation
1) One day before the experiment, remove the CeturegelTM matrix gel from the freezer and place it in the refrigerator at 4°C overnight to melt, and pre-cool the consumables used.
2) Place the CeturegelTM Matrix Gel in an ice box at all times before the experiment.
3) Open the sterilization package of angiogenic slides and remove the slides.
4) Add 10 μl of CeturegelTM Matrix Gel to each well, taking care that the tip of the gun should be perpendicular to the top of the inner well when adding CeturegelTM Matrix Gel to prevent any matrix gel from flowing through the upper well and leaving residual gel.
- Gel
1) First, cover the slide with a lid and prepare a 10 cm Petri dish by placing a water-soaked paper towel to make a wet box.
2) Place the slide into the petri dish and cover the lid.
3) Put the whole petri dish into a CO2 incubator and leave it for about 30 min to wait for the gel to condense, while preparing the cell suspension.
- Cell spreading
1) Prepare the digested cells into a cell suspension with a density of 2×105 cells/ml and mix well.
2) Remove the blood vessel slides containing blood vessels that have solidified into gel.
3) Add 50μl of cell suspension to each well, taking care to keep the tip of the gun perpendicular to the top of the upper well and not touching the gel in the lower well.
4) Add the cell culture medium, cover and let it stand, after a period of time, all the cells will sink down and fall on the surface of the matrix gel.
- Image Acquisition
Observe take photos and keep images regularly according to the cell growth rate.
- Presentation of results
Note：Angiogenesis and fluorescence results
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