Extracellular matrix gel application operation guide
The extracellular matrix gel is a soluble basement membrane preparation extracted from EHS mouse tumor rich in extracellular matrix proteins. Its main components consist of laminin, type IV collagen, heparan sulfate proteoglycan (HSPG), nidogen, and other growth factors such as TGF-beta, EGF, IGF, FGF, tissue plasminogen activator, and other growth factors inherent to EHS tumors. In the mid-1980s, J. Engelbreth-Holm and Richard Swarm from Denmark respectively discovered and specifically described the extracellular matrix gel. The main production process involves extracting/washing the tumor homogenate with saline to obtain soluble proteins, followed by solvent extraction of insoluble complexes from the product, and dialysis with a buffer solution at low temperatures to obtain a colorless to pale yellow solution.
The extracellular matrix gel provides support, tensile strength, and scaffold support for tissues and cells. It also serves as a three-dimensional architecture for cell adhesion and movement, as well as a repository for growth factors, chemotactic factors, and cytokines. Additionally, it can be utilized as a signaling molecule for cellular morphogenesis and differentiation.
Figure: Application Directions of Extracellular Matrix Gel
The following results were obtained from experiments using Arcegel extracellular matrix gel and other accompanying reagents developed and produced by Arcegen Biologicals.
Invasion Assays and 3D Cultures
Angiogenesis
Figure: Results of HUVEC Cell Angiogenesis Assay and Immunofluorescent Staining of Blood Vessels
Intratumoral
Figure. HepG2 cells were subcutaneously implanted into female BALB/c-nu mice and observed for 1 month.
Organoid culture
Related products - Matrix gel
|
Product Types |
Product Name |
Catalog Number |
Specification |
|
|
Basic Concentration
|
C231001 |
5/10 mL |
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|
C231002 |
5/10 mL |
|||
|
Low Growth Factor |
C231003 |
5/10 mL |
||
|
C231004 |
5/10 mL |
|||
|
High Concentration
|
C231005 |
5/10 mL |
||
|
Arcegel Matrix High Concentration, Phenol Red-Free, LDEV-Free |
C231006 |
5/10 mL |
||
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C231007 |
5/10 mL |
|||
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Stem Cell-Specific |
C231008 |
5/10 mL |
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|
Organoid-Specific |
Arcegel Matrix for Organoid culture, Phenol Red-Free, LDEV-Free |
C231009 |
5/10 mL |
|
Related Products - Cytokines
Results of Wnt3a-like Organ Culture Case
|
Protein Name |
Species |
Catalog Number |
Specifications |
|
Wnt3a |
Human |
C230259 |
100/500 ug |
|
R-Spondin 1
|
Human |
C230254 |
100/500 ug |
|
Mouse |
C230257 |
100/500 ug |
|
|
Noggin
|
Human |
C230462 |
100/500 ug |
|
Mouse |
C230612 |
100/500 ug |
|
|
EGF
|
Human |
C230329 |
100/500 ug |
|
Mouse |
C230540 |
100/500 ug |
|
|
FGF-2
|
Human |
C230295 |
100/500 ug |
|
Mouse |
C230522 |
100/500 ug |
|
|
FGF-4 |
Human |
C230346 |
100/500 ug |
|
FGF-7
|
Human |
C230417 |
100/500 ug |
|
Mouse |
C230587 |
100/500 ug |
|
|
FGF-9
|
Human |
C230347 |
100/500 ug |
|
Mouse |
C230548 |
100/500 ug |
|
|
FGF-10
|
Human |
C230418 |
100/500 ug |
|
Mouse |
C230588 |
100/500 ug |
|
|
FGF-19 |
Human |
C230343 |
100/500 ug |
|
Activin A |
Human/Mouse/Rat |
C230520 |
100/500 ug |
|
BMP-2 |
Human/Mouse/Rat |
C230308 |
100/500 ug |
|
BMP-4
|
Human |
C230310 |
100/500 ug |
|
Mouse |
C230249 |
100/500 ug |
|
|
BMP-7 |
Human |
C230311 |
100/500 ug |
|
DKK-1 |
Human |
C230321 |
100/500 ug |
|
HGF |
Human |
C230260 |
100/500 ug |
|
GDNF
|
Human |
C230355 |
100/500 ug |
|
Mouse |
C230552 |
100/500 ug |
|
|
IGF-1 |
Human |
C230371 |
100/500 ug |
|
Shh
|
Human |
C230493 |
100/500 ug |
|
Mouse |
C230624 |
100/500 ug |
|
|
BDNF |
Human |
C230262 |
100/500 ug |
|
NRG1 |
Human |
C230454 |
100/500 ug |
|
SHH
|
Human |
C230493 |
100/500 ug |
|
Mouse |
C230624 |
100/500 ug |
Related Products - Organoid Culture Medium
|
Product Type |
Product Name |
Catalog Number |
Specifications |
|
Mouse Small Intestine |
C231105 |
l00/500mL |
|
|
Colon Cancer (Human) |
C231106 |
l00/500mL |
|
|
Gastric Cancer (Human) |
C231107 |
l00/500mL |
|
|
Liver Cancer (Human) |
C231108 |
l00/500mL |
|
|
Breast Cancer (Human) |
C231109 |
l00/500mL |
|
|
Lung Cancer (Human) |
C231110 |
l00/500mL |
|
|
Esophageal Cancer (Human) |
C231111 |
l00/500mL |
|
|
Ovarian Cancer (Human) |
C231112 |
l00/500mL |
|
|
Pancreatic Cancer (Human) |
C231113 |
l00/500mL |
Other Organoid Culture Reagents
|
Product Type |
Product Name |
Catalog Number |
Specifications |
|
Tissue Cell Digestion
|
C230120 |
100mg |
|
|
C230121 |
100mg |
||
|
C230122 |
100mg |
||
|
C230123 |
100mg |
||
|
C230124 |
100mg |
||
|
Mycoplasma Series |
C230102 |
1mL |
|
|
C230105 |
25/60T |
Appendix
Q:How to melt Arcegel matrix gel, and how long does it take to melt?
A:Place the entire bottle of Arcegel matrix gel in a crushed ice box, then place the ice box in a 4°C refrigerator overnight to melt, ensuring an adequate amount of crushed ice. After melting, rotate the bottle to check if all of the Arcegel matrix gel has melted completely, ensuring no ice crystals or gel chunks remain. We recommend overnight melting, and Arcegel matrix gel with a high protein concentration may require more time. Do not directly place the bottle of Arcegel matrix gel in a 4°C refrigerator to melt, and do not place the ice box and Arcegel matrix gel on the refrigerator door or near the refrigerator door. See the following images for reference:
Q: What is the state of Arcegel matrix gel after melting?
A: Standard Arcegel matrix gel should be clear liquid after melting, with no phenol red, indicating clarity and transparency; the higher the protein concentration, the more viscous the liquid after melting. Please refer to the following image of melted Arcegel matrix gel, with the left two bottles being high-concentration type and the right two bottles being standard type.
Q: How to aliquot and freeze Arcegel matrix gel for use?
A: After thawing, Arcegel matrix gel can be aliquoted into multiple small tubes, all of which should be pre-cooled cryovials, rapidly frozen, and stored to avoid repeated freeze-thaw cycles. All items involved in the process should be pre-cooled before use, including pre-cooled pipettes, tips, and tubes for handling the matrix gel.
Q: What is the reason for finding gel during the melting of Arcegel matrix gel, and why can't it be melted?
A: Arcegel matrix gel is highly temperature-sensitive. It starts to gel above 10°C, and the gelation process accelerates at temperatures exceeding 22°C. If the gelation occurs despite normal conditions upon receipt, it is likely due to improper melting procedures or an increase in environmental temperature during melting, leading to partial gelation. It is crucial to strictly adhere to the melting procedure guidelines and ensure a low-temperature environment during melting.
Q: How to dilute Arcegel matrix gel?
A: Generally, Arcegel matrix gel can be diluted with pre-cooled serum-free culture medium or pH 7.4 PBS. Due to concentration variations between batches, it is recommended to dilute to a specific working concentration rather than using proportional dilutions or dilution factors.
Q: What aspects should be noted when using Arcegel?
A: All consumables and reagents in contact with Arcegel matrix gel should be pre-cooled, and the entire operation should be performed on ice to maintain a low-temperature environment. Arcegel matrix gel is highly temperature-sensitive, and gelation starts once the temperature exceeds 10°C.
Protocol for cytokine usage:
Centrifugation: Upon receipt, the vial contains freeze-dried powder. Prior to opening, quickly centrifuge at 10,000-12,000 rpm for 30 seconds to ensure any potentially dislodged protein adhering to the vial cap or walls settles at the bottom of the vial.
Reconstitution: Reconstitute with the specified solvent as per the instructions, aiming for a concentration range of 0.1-1.0 mg/mL. Note: Avoid vigorous vortexing, as it may cause protein denaturation. Gentle tapping with a pipette tip or allowing the solution to sit at room temperature for a period can aid in dissolution.
Dilution: Dilute with PBS or basal culture medium solution containing 0.1% BSA, 10% FBS, or 5% trehalose (ensure sterility). Use any of the aforementioned diluents to dilute the reconstituted protein solution, adjusting the dilution concentration based on your desired working concentration, but not lower than 10 μg/mL. Aliquot into small tubes and store at -20 to -80°C.
Usage: When needed, thaw one aliquot of the cytokine and add it to your culture medium to achieve the desired concentration. Avoid repeated freeze-thaw cycles for cytokines.