Extracellular Matrices and Basement Membrane Extracts
Substrates that mimic the extracellular matrix (ECM) in culture have been shown to be imperative for robust cell growth, optimal health, and as a method to direct the behavior and structural formation of cells in vitro. Basement membrane extract (BME) is a commonly used scaffolding for 2- and 3-D cell culture, including pluripotent stem cell expansion and differentiation as well as spheroid and organoid formation. BMEs contain laminin, fibronectin, collagen IV, vitronectin, entactin, and heparan sulfate proteoglycans. This mixture of ECM proteins within the hydrogel plays an important role in directing cell behavior and health, providing not only a critical growth substrate but also an optimized environment for cell-to-cell interactions to facilitate in vivo-like growth dynamics. While BME is being utilized across a wide breath of applications, the quality and behavior of cells cultured in BME can greatly be impacted by handling, hydrogel concentration, and coating methods.
Choose the Best Cell Matrices for Your Research
Cultrex matrices are carefully formulated to mimic the complex composition and physical properties of the native ECM, providing a more physiologically relevant environment for your cells to grow and function in. By using Cultrex, you can more accurately replicate the in vivo microenvironment and achieve more reliable and meaningful results. Cultrex is rigorously tested for quality and consistency, ensuring that you get the same reliable results every time.
As a leading brand in the development and production of high-quality cell matrices, our extensive range of Cultrex ECM products is designed to meet the specific needs of a diverse range of fields, including cancer biology, stem cell research, and tissue engineering.
See the selection guide below to purchase today or reach out to a specialist to discuss how we can provide a reliable source of ECM for your research or help you find the right matrix.
Cultrex BME Matrix Selection Guide
|Application||Product||Without Phenol Red||With Phenol Red||Matrigel Equivalent|
|Stem Cell, Organoid, 3-D Culture||Cultrex UltiMatrix||BME001-05 (RGF)||-||356255, 354248, 354262, 354263|
Cultrex Basement Membrane Extract, Type 1
356234, 354234, 356237, 356232, 356235, 356254
|3433-010-01 (RGF)||3433-010-01P (RGF)||356230, 354230, 356231, 356252, 356238, 356253, 356239|
Cultrex Basement Membrane Extract, Type 2
|3533-010-02 (RGF)||3533-010-02P (RGF)|
|3536-005-02 (RGF, Select)||-||-|
|Difficult to Culture Organoids||Cultrex Basement Membrane Extract, Type R1||3433-010-R1||-||356255|
|Xenograft, Tumorgraft||Cultrex Basement Membrane Extract, Type 3||3632-010-02||3632-010-02P||-|
Cultrex Stem Cell Qualified Basement Membrane Extract
354277, 356277, 356278
|3434-050-RTU (RGF, ReadyBME)||-|
|3-D Culture||Cultrex 3-D Culture Matrix||3445-010-01 (RGF)||-||-|
Cultrex Extracellular Matrix Proteins
Cultrex ECM Proteins provide a defined substrate for cell adhesion in culture. Many proteins of the ECM interact with cells via cell surface integrin family receptors. The resulting focal contacts are important for the maintenance of tissue architecture and for supporting a variety of cellular processes. ECM molecule/integrin binding may initiate a complex network of signal transduction cascades that, depending on the context, play an important role in cell spreading, migration, proliferation, and differentiation during embryogenesis, wound healing, and tumor development. To accommodate a variety of different models, we have developed a full range of ECM component proteins including Laminin I, Collagen I, Collagen IV, Fibronectin, and Vitronectin. These include purified proteins that have been qualified for 3-D or stem cell cultures.
|Mouse Laminin I||A purified basement membrane protein that has been developed, produced and qualified specifically for cell culture studies where the presence of pathogens may cause concern.|
|Mouse Collagen IV||Used for coating of tissue culture surfaces to promote cell attachment and proliferation and to study its effects on cell behavior.|
|Rat Collagen I||Major structural component of extracellular matrices found in connective tissue and internal organs, used to promote cell attachment, growth, differentiation, migration, and tissue morphogenesis during development. The 5 mg/ml concentration is very viscous for researchers wanting a thicker Collagen.|
|Rat Collagen I, Lower Viscosity||Diluted to a lower concentration (3 mg/ml); it is less viscous and easier to handle. A major structural component of extracellular matrices found in connective tissue and internal organs, but is most prevalent in dermis, tendons, and bone. It promotes cell attachment, proliferation, differentiation, migration, and tissue morphogenesis.|
|Vitronectin||Vitronectin is a large glycoprotein found in blood and the ECM. Vitronectin contains multiple binding sites for a variety of structures.|
|3-D Culture Extract Laminin I||3-D Culture Matrix™ Mouse Laminin I may be used as a gel on which to grow cells, as a media additive alone or in concert with other basement membrane components, or to study cellular growth and differentiation in three dimensions in vitro.|
|3-D Culture Extract Rat Collagen I||The 3-D Culture Matrix™ Collagen I may be used as a gel on which to grow cells or a media additive alone or in concert with other basement membrane components to study cellular growth and differentiation in three dimensions in vitro.|
Best Practices for Handling Cell Matrices
- Keep all stock solutions at -80 °C for long term storage (≥ 1 month)
- Note: Cultrex ReadyBME is pre-diluted and should be stored at 2-8 °C, never frozen
- Thaw frozen Cultrex BME matrix on ice in a 2-8 °C refrigerator overnight. This allows for ample thawing while maintaining the product at a stable cold temperature
- Aliquot into working amounts. Store aliquots at -80 °C for long term storage
- Avoid storing Cultrex BME matrices at 2-8 °C for > 1 day
- Cultrex BME matrices will gel if temperature is ≥ 18 °C, so work quickly and always on ice and with pre-chilled supplies and tools, such as pipette tips
- Always dilute Cultrex BME matrices with ice-cold cell culture media
This video provides best practices and protocols for working with Cultrex BME, including thawing, aliquoting, thin layer method, thick layer method, and dome formation. R&D Systems’ Cultrex BME portfolio provides consistent and soluble extracellular matrices used globally for cell culture applications that require attachment or a three dimensional matrix for complex tissue development. Cultrex BME matrices are available for a variety of in vitro applications including pluripotent stem cell, spheroid, and organoid culture. Techniques in this video apply generally when using Cultrex BME. Cultrex BME should be maintained at -80 °C for long-term storage. To thaw Cultrex BME, remove a vial from the -80 °C freezer, submerge it in ice, and place it in a refrigerator overnight. We recommend placing pipettes, cryovials, and other materials needed to aliquot or dilute Cultrex BME in a -20 °C freezer so that they are chilled prior to use. The next morning, remove the matrix from the refrigerator while keeping it on ice. Gently swirl the matrix on ice to ensure a homogeneous mixture before aliquoting or downstream use.
Aliquoting Cultrex BME will extend the lifetime and preserve the quality of the matrix by limiting freeze-thaw cycles that compromise matrix composition and polymerization. First, place the thawed Cultrex BME (submerged in ice) in a cell culture hood. Fluctuations or temporary exposure of Cultrex BME to temperatures above 2-8 °C can cause changes in protein polymerization that could impact the matrix consistency or cell behavior. To ensure the matrix stays cold, we recommend chilling all pipettes and pre-labeled vials in the -20 °C freezer prior to aliquoting. Mix Cultrex BME by pipetting up and down to achieve a homogeneous solution. Aliquot the matrix at a dilution recommended by the manufacturer or that suits your desired application. Use reverse pipetting techniques to minimize bubble formation when pipetting Cultrex BME. You can store aliquots of Cultrex BME at -80 °C for up to six months.
The thin coating method is commonly used for the monolayer culture of adherent cells as well as to expand embryonic and induced pluripotent stem cells. We recommend Cultrex Ultimatrix BME or Stem Cell Qualified Reduced Growth Factor BME for thin coating. Using pre-chilled pipette tips, mix thawed Cultrex BME by pipetting the solution up and down. Avoid introducing bubbles during this process. Dilute Cultrex BME to the desired concentration in cold serum-free medium. Optimal coating concentration may vary by cell type or application and should be empirically determined. Add dilute matrix to the vessel for coating. Sufficient volume should be added to cover the growth surface of the vessel. Swirl the vessel to encourage full surface coating. Incubate the coated object for one hour at room temperature or at 37 °C for 30 to 45 minutes. Incubation at 4 °C overnight results in a thicker coating, which can be beneficial for some cell lines. Once solidified, aspirate the remaining solution. It is important to make sure the Cultrex BME coating does not dry out, so move on to the next step immediately or seal the plate with parafilm until you are ready to culture.
The thick layer method uses undiluted Cultrex BME to promote the formation of 3-D structures including spheroids. Cells can be plated on top of the concentrated matrix or embedded within the matrix. Using cooled pipette tips, mix thawed Cultrex BME by pipetting the solution up and down. Avoid introducing bubbles during this process. To create a thick layer for growing cells on, either dispense Cultrex BME directly into the culture vessel or if you are embedding cells in the matrix, mix the cell suspension with Cultrex BME prior to adding into the culture vessel. Add enough volume to cover the growth surface. Incubate the plate at 37 °C for 30 minutes to solidify the matrix. Once solidified, aspirate any unpolymerized material and rinse with serum-free medium. Add the appropriate cell culture media to each well, while being careful not to disrupt the matrix surface.
Cultrex Ultimatrix BME and Organoid Qualified Cultrex Reduced Growth Factor BME Type 2 are used to generate domes for organoid culture. Prepare your cell culture vessel by pre-incubating it in the 37 °C incubator. The pre-warmed plate promotes proper dome formation when plating. Using cooled pipette tips, mix thawed Organoid Qualified Cultrex BME or Cultrex Ultimatrix BME by pipetting the solution up and down. Avoid introducing bubbles during this process. Dilute the organoid solution directly in ice-cold Cultrex BME and mix thoroughly. A final matrix concentration of greater than eight milligrams per milliliter is recommended to ensure dome formation. Slowly dispense organoid and matrix mixture onto the center of the pre-warmed culture dish. Use reverse pipetting techniques to minimize bubble formation when creating domes. Depending on your organoids, domes can be arranged as a single dome per well or six to eight domes per well. Incubate the plate at 37 °C for 30 minutes to solidify the matrix. Inverting the plate while incubating may help retain dome formation and prevent settling of organoids on the plates. Once the domes are polymerized, add growth media into the culture wells, being careful not to disturb the Cultrex BME domes. Your Cultrex BME domes are ready for continued culture.
Basement membrane hydrogels have unique physical properties. They are in a liquid state at 2-8 °C and solidify when brought to room temperature. While a liquid, Cultrex BME matrices can be easily manipulated and diluted, but any fluctuations or temporary exposure to temperatures above 2-8 °C can potentially impact the health and behavior of the cell culture system.
When working with Cultrex BME matrices or other ECMs, it is recommended to use chilled pipettes, tubes, and other tools. Additionally, it is crucial to avoid vigorous shaking or vortexing, as this can cause the matrix to solidify prematurely or lead to the formation of air bubbles, which can interfere with cell culture.
Cultrex BME Matrices should be stored at -20 °C or below, in their original container, and used before the expiration date. Avoid repeated freeze-thaw cycles, as this can damage the physical properties of the matrix and affect its performance.
Select the Optimal Coating Method for Your Cell Culture
Cultrex BME is a soluble form of basement membrane that can be applied to a variety of cell culture conditions. However, it is important to understand the various coating methods that can be employed and the impact they can have on your cell culture. The method of use is dictated by multiple factors, including the type of starting material (single cells or tissue), the adherence requirements of your cell or tissue type, and the need for a 2- or 3-D culture system.
|Thin Coat||Cells are grown on top of a thin layer of Cultrex BME||Primary Cell Propagation
Cell Invasion Assays
|Thick Coat||Cells are grown on top of a thick layer of Cultrex BME||
|Sandwich||Cells are cultured between two thick layers of Cultrex BME||
|Embedded-Layer||Cells are cultured while embedded in Cultrex BME||
|Embedded-Dome||Cells are cultured while embedded in Cultrex BME and plated into cell culture vessel as domed structures||Organoid Culture|
The thin coating method for Cultrex BME provides an adherent ECM substrate for cell proliferation and maintenance. It is commonly used for the expansion of embryonic and induced pluripotent stem cells but can be broadly applied to cell lines or primary cells. Benefits of this coating method are its simplicity to set up and its accessibility for immunocytochemistry and cell imaging downstream.
The sandwich coating method for Cultrex BME provides a more complex ECM microenvironment for cell growth and differentiation in a more in vivo-like 3-D environment. It is also beneficial for cultures where a larger adherence area is needed to accommodate mechanical stress. While this method takes slightly more work than the thin coating method, the cells are fully embedded into the matrix, providing a more rigid 3-D scaffolding environment.