Extracellular matrix gel application operation guide

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

 

Arcegel Matrix LDEV-Free 

C231001

5/10 mL

Arcegel Matrix Phenol Red-Free, LDEV-Free

C231002

5/10 mL

Low Growth Factor

Arcegel Matrix GFR, LDEV-Free Matrigel

C231003

5/10 mL

Arcegel Matrix GFR, Phenol Red-Free, LDEV-Free

C231004

5/10 mL

 

 

High Concentration

 

Arcegel Matrix High Concentration, LDEV-Free

C231005

5/10 mL

Arcegel Matrix High Concentration, Phenol Red-Free, LDEV-Free

C231006

5/10 mL

Arcegel Matrix High Concentration, GFR, LDEV-Free

C231007

5/10 mL

Stem Cell-Specific

Arcegel Matrix hESC-Qualified, LDEV-Free

C231008

5/10 mL

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

3DCultr Intestinal Organoid Growth Medium (Mouse)

C231105

l00/500mL

Colon Cancer (Human)

3DCultr Intestinal Cancer Organoid Growth Medium (Human)

C231106

l00/500mL

Gastric Cancer (Human)

3DCultr Gastric Cancer Organoid Growth Medium (Human)

C231107

l00/500mL

Liver Cancer (Human)

3DCultr Liver Cancer Organoid Growth Medium (Human)

C231108

l00/500mL

Breast Cancer (Human)

3DCultr Breast Cancer Organoid Growth Medium (Human)

C231109

l00/500mL

Lung Cancer (Human)

3DCultr Lung Cancer Organoid Growth Medium (Human)

C231110

l00/500mL

Esophageal Cancer (Human)

3DCultr Esophageal  Cancer Organoid Growth Medium (Human)

C231111

l00/500mL

Ovarian Cancer (Human)

3DCultr Ovarian cancer Organoid Growth Medium (Human)

C231112

l00/500mL

Pancreatic Cancer (Human)

3DCultr Pancreatic cancer Organoid Growth Medium (Human)

C231113

l00/500mL

 Other Organoid Culture Reagents

Product Type           

Product Name   

Catalog Number

Specifications

 

 

Tissue Cell Digestion

 

Collagenase I 

C230120

100mg

Collagenase II 

C230121

100mg

Collagenase III 

C230122

100mg

Collagenase IV

C230123

100mg

Collagenase V

C230124

100mg

 

Mycoplasma Series

DfCell 1000×Mycoplasma Prevention Reagent

C230102

1mL

DfCell Mycoplasma LAMP Detection Kit

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.


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