NEST Biotechnology Co., Ltd. (NEST)
NEST - Model GelNest™ Matrix -Mouse Tumor Tissue and Contains Extracellular Matrix Component
GelNest™ Matrix is derived from mouse tumor tissue and contains extracellular matrix components such as laminin, type IV collagen, heparan sulfate proteoglycans, and more. These components support cell adhesion, differentiation, and proliferation, providing signals for these processes. Additionally, they simulate the characteristics of the basement membrane in the physiological environment, enhancing the success rate and effectiveness of cell culture. In addition to the matrix components, GelNest™ Matrix is rich in various growth factors. These growth factors promote cell differentiation, proliferation, and migration, mimicking cellular signaling pathways and interactions in the physiological environment. GelNest™ Matrix has a wide range of applications, particularly in tissue engineering, cell culture, and research. It can be used for organoid culture, stem cell differetiation, angiogenesis, migration or invasion assays, and in vivo tumor studies.
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Organoid Culture Test
- Re-suspend the single-cell suspension for organoid culture in pre-cooled basal medium at 4°C and perform cell counting.
- Mix the cells with the gel solution and add the mixture to pre-warmed 24-well plates. Each well should contain approximately 5x104 cells and 60µL of matrix gel.
- Immediately place the plates in the incubator, and the gel will solidify in approximately 10 minutes.
- Add 500µL of organoid culture medium for cultivation.
- Wait for 3-5 days for the organoids to form. Finally, image the live cells using high-content microscopy to determine the sensitivity of the organoids to various drugs.
This method provides an efficient and convenient solution for organoid culture and can be used in areas such as drug screening and tumor research.
Stem Cell Differentiation Test
For human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) without feeder layer culture:
- Thaw GelNest™ Matrix stored in freezing conditions and let it thaw overnight in a 4°C ice bath. Gently blow and mix the gel three times with a pre-cooled pipette tip for thorough mixing. Transfer the thawed matrix gel using the pre-cooled pipette tip. If bubbles form, briefly centrifuge with a handheld centrifuge to remove them.
- Preheat the cell culture plates in the incubator.
- Dilute the gel solution in pre-cooled serum-free medium at a 1:100 ratio. Make sure to cover the entire culture plate with the diluted gel solution. The recommended amount is 300µL/cm^2 in the culture dish.
- Leave the culture plates with the modification solution at room temperature for 1 hour.
- Remove the modification solution and immediately seed the stem cells mixed with mTeSR in the culture plate. Be cautious to prevent the surface of the modified culture plate from drying out.
This method provides an efficient and convenient solution for stem cell culture and is expected to play an important role in tissue engineering, regenerative medicine, and other fields.
In Vitro Blood Vessel Formation Test
- Replace the complete growth medium with starvation medium for cells: DMEM medium containing 0.2% FBS, 2mM L-glutamine, 1mM sodium pyruvate, 100U/mL penicillin, and 100µg/mL streptomycin. Starve the cells for 24 hours.
- Spread 50µL of GelNest™ Matrix evenly on the bottom of a 96-well plate. To prevent the gel from adhering to the pipette tip, aspirate and blow FBS once in the pipette tip to wash the inner wall of the pipette tip with FBS.
- Place the 96-well plate in a 37°C cell culture incubator and incubate for 30 minutes to solidify the gel.
- Digest the endothelial cells and perform cell counting.
- Add 5x10^4 HUVEC cells to the 96-well plate containing the gel, totaling 200µL of cell suspension. Place the 96-well plate in the incubator for cultivation.
- Vascular-like network structures will form within 3 to 12 hours. This is the optimal observation time.
- At the optimal observation time, carefully remove the culture medium and stain with culture medium containing a 1/1000 concentration of Calcein AM (green). Image the cells using a microscope and record the morphology and characteristics of the vascular network.