Traditional cell culture involves growth in a monolayer where cells are flat and can adhere and proliferate freely on a horizontal plane but do not support diffusion in the vertical direction. Cells cultured on 2D surfaces exhibit strong apical-basal polarity, which is associated with certain cell types, such as epithelial cells. 3D cell culture, also known as three-dimensional cell culture, refers to the co-culture of various cells with carriers made of materials possessing three-dimensional structures in vitro, allowing cells to migrate and grow in the three-dimensional spatial structure of the carrier, forming a three-dimensional cell-carrier complex. This method is mainly used for culturing cell spheroids, organoids, and similar structures.

Figure 1. Difference between ECM-coated glass or plastic surface (2D) and typical 3D ECM [1]

 

As a disease validation model, organoids cultured in a 3D environment provide a closer approximation to the in vivo conditions compared to traditional 2D disease models, elucidating disease progression, homeostasis, and pathogenesis. They are poised to play a crucial role in various fields such as cell therapy, drug development, genetic engineering, immunology, tissue regeneration, etc. The matrix gel, as an important carrier for organoids and 3D culture, plays a pivotal role in these applications.

 

Table 1. Comparison between 2D and 3D Cultures [1]

Arcegel Matrix Gel is a soluble basement membrane preparation extracted from EHS mouse tumors rich in extracellular matrix proteins. Its main components consist of laminin, type IV collagen, heparan sulfate proteoglycans (HSPG), nidogen, and other growth factors such as TGF-beta, EGF, IGF, FGF, tissue plasminogen activator, and other growth factors present in EHS tumors. It provides support, tensile strength, and scaffold support for tissues and cells, serves as a three-dimensional architecture for cell adhesion and movement, and acts as a reservoir for growth factors, chemokines, and cytokines. It also serves as a signal for cell morphogenesis and differentiation. Arcegel Matrix Gel, developed and produced by Arcegen Biology, is free from LDEV (lactate dehydrogenase-elevating virus), has an ultra-low endotoxin content, and has been thoroughly tested for mycoplasma to ensure no contamination, including different types of matrix gels such as basic concentration, high concentration, and low growth factor concentration.

 

Application directions

Product Features

High Safety: Free from LDEV (Lactate Dehydrogenase-Elevating Virus)

Diverse Concentrations: Concentration ranges between 8 to 20 mg/mL

Good Batch Stability: Stringent production quality inspection processes ensure stable performance between batches

Low Endotoxin Levels: Endotoxin content ≤ 4 EU/mL

Contaminant Detection: Tested free of mycoplasma, bacteria, and fungal residues

High Batch Yield: Batch yield surpasses the L-level

Excellent Compatibility: Compatible with any type of cell culture medium

 

Validation for 3D Cultures

 

Preparation of Cell Suspension

Pre-chill the 24-well cell culture plate and pipette tips on ice. Place the matrix gel on ice or at 4°C to thaw slowly. Retrieve logarithmic-phase HepG2 cells and gently rinse with PBS, followed by digestion with trypsin. After digestion is stopped, prepare and collect single-cell suspension. Centrifuge at 1500 rpm for 3 minutes, discard the supernatant, and resuspend the cells in DMEM complete medium. Count the cells using a hemocytometer and adjust the cell concentration to 1.5×10⁵ cells/mL.

Cell Seeding

Mix the thawed matrix gel and adjusted single-cell suspension at a 1:1 ratio on ice, then gently pipette 40~50μL of the mixed single-cell suspension vertically into pre-chilled 24-well plates to form arched cell droplets.

Results statistics

After stabilizing in a cell culture incubator at 37°C with 5% CO2 saturation for 30 minutes, add 1~2mL of DMEM complete medium to each well and continue incubating. Observe and photograph daily.

Figure 2. Results of Cell 3D Culture after 4 Days

 

FAQ

01. What causes the color change (from pale yellow to deep red) in the obtained matrix?

For matrix gel containing phenol red, the main reason for the color change is the reaction between phenol red and bicarbonate with CO2. However, the color difference will decrease after balancing with 5% CO2. After thawing, gently shake the reagent bottle to evenly disperse the matrix gel.

02. What precautions should be taken when handling the matrix gel?

All operations should be performed under aseptic conditions, and pre-chilled pipettes should be used to ensure that the matrix gel is evenly dispersed.

03. How to aliquot and freeze the matrix gel for later use?

After thawing, the Arcegel matrix gel can be aliquoted into multiple tubes. All aliquoting should be done using pre-chilled storage tubes, rapidly frozen, and stored to avoid repeated freeze-thaw cycles. All items involved in use should be pre-chilled, and matrix gel handling should be done using pre-chilled pipettes, tips, and tubes.

 

Related products

Product type	Product Name	Catalog Number	Specifications
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