Generic Formulation - NON-Degradable WITHOUT RGD peptides - in vitro Assay Kit
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Generic Formulation - NON-Degradable WITHOUT RGD peptides - in vitro Assay Kit Generic Formulation - NON-Degradable WITHOUT RGD peptides - in vitro Assay Kit Generic Formulation - NON-Degradable WITHOUT RGD peptides - in vitro Assay Kit Generic Formulation - NON-Degradable WITHOUT RGD peptides - in vitro Assay Kit Generic Formulation - NON-Degradable WITHOUT RGD peptides - in vitro Assay Kit

Generic Formulation - NON-Degradable WITHOUT RGD peptides - in vitro Assay Kit

QGel® GENERIC Formulation NS73-A adapts to many cell types.

No- MMP Degradable

No- RGD Peptide, cell adhesion properties

The 3D Matrix formulaiton is Non-MMP Degradable and contains NO RGD peptides for cell adhesion.  This formulation well-suited for research applications involving many cancer cell lines.  The Assay Kit format is ideal for high throughput screening. 

Encapsulation Video 

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The QGel® Assay Kit has been developed for high-throughput cell-based assays and is compatible with existing standard liquid-handling robots.  Each kit comes with protocols tailored to your research needs. Standard cell suspension is added to the Assay Kit to resuspend the lyophilized powder gel formulation and the combined mixture of cell suspension and QGel Matrix is then further dispensed into the separate assay plates.  Most of our clients work with 384-well assay plates, but we are able to help you establish your own protocols for 96-well or 1536-well assay plates as well.

 

This video demonstrates how cells are encapsulated with QGel Assay Kits.

 

FEATURE BENEFITS OF QGEL MICROENVIRONMENT SPECIFIC MATRICES

 

Fully Synthetic

Each hydrogel-based QGel Matrix is developed by modulating various properties of the extra-cellular microenvironment (ECM) including cell adhesion-promoting ligands, matrix degradation kinetics initiated by natural cell secretion and ECM stiffness characteristics. The result is a unique synthetic ECM designed to recapitulate in vivo-like cell behavior and tissue growth. This formulation is free of growth factors or any cell culture media ingredients.

 

Reproducible

Because QGel ECM Matrices are developed with fully synthetic bioactive components, there is no variability from batch to batch and there is no contamination risk.  This means you can focus your research efforts to achieve the desired endpoints of your research study.

 

A Complete Discovery Solution:

Incorporating the full line of QGel products in your research means that your assets can move upstream and downstream without the need to discover new platform technologies at each step of the way.

integrated_solutions 

In Vitro to In Vivo Research

Each in vitro QGel formulation has a companion formulation developed for in vivo animal studies.  QGel Matrices offer highly effective solutions for achieving even the most difficult to grow tumor xenograft development in animals.

 

Easy Handling

Easy to follow protocols are included with each QGel Assay Kit and our dedicated lab technicians provide the assistance necessary to ensure the assays are developed correctly at your labs.   

QGel Assay Kits work with any standard liquid-handling robot.  Simply add the cell suspension with the lyophilized powder in the QGel Assay Kit, resuspend and dispense into your assay plates. The hydrogel matrix will form within minutes under normal temperature conditions with the cells encapsulated. 

Click here to view a video of encapsulating cells with the QGel Assay Kits.

Click here to view a video on drug screening with QGel Matrices at various timepoints after cell encapsulation.

 

High Content Data Analysis

Image Analysis

  • -Number of spheroids
  • -Spheroid size distribution
  • -Spheroid morphology
  • -Immunofluorescence

Biomarker

  • -Protein expression
  • -RNA & DNA expression

Proliferation

  • -Metabolic activity
  • -DNA

 

Storage

QGel has a shelf life of 5 years. Store at -20C.

Organ -Non organ-specific
Disease Cancer
Platform In Vitro
Use Automated, Robotic
Optimized for Cell Line -Generic / Non-cell specific
Gel Volume 100 µL per mini vial
Product Type QGel 3D Matrix Products

Included with each purchase of a QGel Assay Kit is service customer support for translating QGel Assay Kits into your workflow.  Below are the most common protocols needed for this product.  Please do not hesitate to contact us if you require assistance. 

 

PROTOCOLS

T.1.A Cell Encapsulation and Culture- 384-well format

PDF Download: Cell Encapsulation and Culture protocol - 384-well plate format

Video:

 

T.1.B Cell Encapsulation and Culture: Media Addition- 384-well format

PDF Download: Cell Encapsulation and Culture protocol - 384-well plate format

Video:

 

T.1.C Cell Encapsulation and Culture: Media Change- 384-well format

PDF Download: Cell Encapsulation and Culture protocol - 384-well plate format

Video:

 

T.2.1.A Cell Proliferation AlamarBlue® Assay- 384-well format

PDF Download: Cell Proliferation AlamarBlue® protocol - 384-well plate format

Video:

 

T.2.1.B Cell Proliferation AlamarBlue® Assay: Gel Wash- 384-well format

PDF Download: Cell Proliferation AlamarBlue® protocol - 384-well plate format

Video:

 

T.2.2.A Calcein AM Cell Viability Assay- 384-well format

PDF Download: Calcein AM Cell Viability Assay protocol - 384-well plate format

Video:

 

T.2.2.B Calcein AM Cell Viability Assay: Gel Wash- 384-well format

PDF Download: Calcein AM Cell Viability Assay protocol - 384-well plate format

Video:

T.3.1 Cell Fixation for Staining- 384-well format

PDF Download: Cell Fixation for Staining - 384-well plate format

We offer a $200 discount voucher to clients who: (1) use the QGel ECM Discovery Kit to identify matrices for use with new cell lines not found in our existing cell line library, and (2) share that data with us for use on our website for the benefit of other QGel users.  A $200 discount voucher will be given for each cell line, up to 10 cell lines per calendar year upon the acceptance of the data submission form found below. This voucher may be used on future purchases and is valid for 2 years.

To Qualify for this voucher, email us webshop@qgelbio.com with the following information:

1.   Cell line profiled
2.   Method used to profile the cell line (Alamar blue, etc)
3.   Cell density per well
4.   QGel Formulation ID used
5.   Media conditions
6.   Image of cell line growth chart
7.   2 images of cell growth taken at various time points
8.   How you would like to be credited as the source of this data, and
9.   Your acceptance of the Consent and Acknowledgement of this program contained below.

If we chose to use your data on our website, you will be credited with a $200 coupon/voucher (per cell line) for use towards your next purchase.  You will be notified regarding our acceptance of your data shortly after your submission along with the terms and conditions.

Please contact us if you have any questions about this program.

Consent and Acknowledgement

I hereby assign QGel the right to use this data on the www.qgelbio.com website. However, I retain full ownership of this data. QGel may not use this data anywhere else but on the www.qgelbio.com website. I certify that the data submitted is accurate to the best of my knowledge and was obtained using quality scientific methods. I grant QGel the right to include me/my lab as the source of this information as entered above. This information is provided for the informational benefit of users of the qgelbio.com website and I make no warranties as to its accuracy and submit this data as is. I grant no guarantee of the accuracy of this data and can not be held liable for any reason in the event that other users obtain results that differ from the data I have submitted.

Frequently Asked Questions

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What is QGel made of?

QGel Matrices are PEG (polyethylen glycol) based material. This polymer is crosslinked into a 3D network with peptides that contain an amino acid sequence that can be cleaved by cell-secreted proteases such as MMPs. Therefore, like in natural matrices, cells are able to degrade the matrix locally via secretion and activation of proteases (MMPs), move further and proliferate. Furthermore, if needed, QGel provides gels with covalently-bond RGD peptides to stimulate cell-integrin binding and allow adhesion of the cells to their environment.

How do I know which QGel Matrix product fits to my experimental needs?

Based on years of research and feedback from clients, we have screened a multitude of varying microenvironmental factors (including adhesion ligands, degradation kinetics and matrix stiffness properties) to identify formulations that tend to favor specific cell types. You can search for cell specific gel formulations in the product filter block in the webshop catalog. We also provide Microenvironment Screening Kits if you are unable to identify formulations for your specific cell type. These kits provide 4 different formulations with different properties for you to identify the microenvironment that best suits your needs. Once you have identified the formulation that best supports your research objectives, future orders can be placed for that specific formulation. We provide discount incentives if you share with us the formulations that best supported your specific cell type. Contact us or view your MY ACCOUNT in this webshop and click on the REFERRALS TAB to learn about our discount program.

What is the degradation rate of the MMP-degradable QGel Matrix?

The degradation kinetics of the gels depends on the ability of the cells encapsulated in the gels to produce/activate MMPs. Without this cell-mediated proteolytic activity the gels remain intact and do not degrade. Degradation components (e.g. MMP sensitive or insensitive peptides) act as crosslinks of the PEG molecules and determine the MMP sensitivity of the gels. These amino acid substrates with different MMP-sensitivities were characterized using MMP-1 (Lutolf, PNAS 2003). The MMP sensitive substrate was also degraded by other MMPs (Lutolf, Nature Biotechnology, 2003). In contrast the MMP-insensitive substrate was shown to be quite stable (i.e. essentially non-degradable) in vitro and in vivo experiments (Lutolf, PNAS 2003) (Rizzi, Biomacromolecules, 2006).

What is the cell viability in QGel Matrices?

Cell viability in QGel Matrices is very high. For example, it was shown that viability of human foreskin fibroblasts was around 90-95% 16 hours after encapsulation (Raeber et al., Biophysical Journal 2005). These cells were seen to form 3D cellular networks within the gels after some weeks of cell culture, demonstrating potentiality for long term cell culture. However, note that cell viability rate in 3D gels depends on the cell type used.

What type of cells can be cultured successfully in QGel Matrices?

QGel Matrices can be used to encapsulate most cell types. This includes cell lines as well as primary cells. Please contact us directly if you are looking for matrices that support primary cell growth or are interested in developing organoid models.

What is the stiffness of QGel Matrices?

Shear modulus (G’) of the soft QGel matrices range from 800-1000 Pa. Contact us directly for more information regarding the stiffness of your specific QGel formulation.

Do QGel Matrices swell or shrink after cross-linking reaction?

Please contact us directly to verify the amount of swelling to expect with your particular QGel formulation. We can provide the exact swelling data in the certificate of analysis that are generated with the manufacturing of each QGel Matrix. This data is without encapsulated cells. If cells are encapsulated, swelling could vary depending on the cell seeding density. This aspect requires optimization by the end user depending on the design of the research study. Please also take note that swelling may be slightly different within the product range of gels.

Are the mechanical properties and the swelling ratio the same for every QGel Matrix product?

No, they will vary depending on the stiffness properties of each unique gel formulation. However, this has rarely been considered an issue. Please contact us if you think swelling might be an issue and we can let you know how much swelling to anticipate with your specific QGel formulation.

How can I incorporate bioactive molecules in QGel Matrices?

Bioactive molecules can be incorporated into QGel Matrices during gelation. However, the strategy of incorporation and delivery strongly depends on the nature of the molecule, and this aspect requires further investigation by the end user. Examples of incorporation of peptides, BMP and VEGF can be found in published works. Do not hesitate to contact us for these published works or if you would like help to design your experiment to account for the addition of bioactive moleculs during gelation.

What is the concentration of QGel Matrices?

In general, the range of QGel Matrices have similar mechanical characteristics (e.g. shear modulus G’) and that differ exclusively in the biological and biochemical properties that characterize each product. Shear modulus (G’) of the QGel Matrices are in the range of 800-1000 Pa. Contact us regarding the specific mechanical characteristics of your selected QGel formulation.

What is the protocol for cell encapsulation?

Please review the Protocol tab included on this product description page.

What are the recommended gel disc dimensions for in vitro experiments?

The recommended volume of the gel discs depends on your experimental needs, e.g. number of gels and required number of cells per gel in order to perform the different investigations planned (conditions, replicates, ...), as well as the time required to complete the casting of the discs before liquid solution gels. We recommend making gel discs of 30uL to 60 uL. The resulting gel discs yield a diameter of 5 to 8.25 mm and a height of 1.5 mm (which is approximately the height of the QGel Disc Caster). After incubation in liquid, such as cell culture media, the gel discs will swell slightly. For more information on swelling properties contact us. With this disc size, using QGel Matrix (vials of 500uL gel solution each) and the QGel Disc Caster, you can cast between 8 to 16 discs. Detailed instructions on the gel disc casting method can be found here: https://www.qgelbio.com/files/protocols/Casting3D_Discs.pdf It is recommended that you perform some preliminary tests to define the best conditions for your experimental needs.

How many gel discs can be made with one vial of QGel Matrix?

This depends on the volume of each gel disc. We recommend 30 uL to 60 uL volume per gel disc. This results in ca. 16 to 8 gels per Matrix vial (total volume 500 uL). Detailed instructions on gel disc casting can be found here: https://www.qgelbio.com/files/protocols/Casting3D_Discs.pdf

What is the recommended cell density in the gels?

The cell density needs to be optimized for each cell type and experimental condition. Indeed, cell density is a critical factor for cell survival and proliferation. You can start based on cell density used in your previous 3D studies with these cells if good results were obtained. We consider 1?105 cells/mL gel as low cell density and 10?106 cells/mL gel as very high cell density. For more information, please contact us to get an idea of the different cell densities used with QGel Matrices in the past.

Isn't the 500 uL gel solution too much for one experiment?

From one QGel Matrix vial, 500 uL of gel can be made and this is usually the minimum material required to perform an experiment. One experiment involving different conditions with several time points and analysis including replicates may easliy require 500 uL gel.

Could I use a portion of the QGel Matrix vial and freeze the remaining portion?

No, it is NOT possible to to freeze the remaining QGel Matrices. Each QGel Matrix vial can be used only once because hydrogel formation starts as soon as the powder is resuspended into the QGel Buffer. Freezing and thawing processes will alter the properties of the gel. Note that 500 uL of gel can be made from one QGel Matrix vial and this is usually the minimum material required to perform an experiment. One experiment involving different conditions with several time points and analysis including also replicates may require easily 500 uL gel.

How can I increase or decrease the gelation time of QGel Matrices?

QGel provides QGel Buffer A (0.3M HEPES solution) with a pH of 7.9 ± 0.1. At this pH working time with the liquid solution before a gel starts forming is 5-10 minutes. Indeed, the reaction kinetics is pH-dependent: the higher the pH, the faster gelation occurs. The exact gelation time depends on the experimental conditions. If your experiments require longer or shorter gelation times, you can make up your own reaction buffer by altering the pH (higher pH for faster, lower pH for slower). pH sensitivity of the gelation can be seen in the following publication: Lutolf M, Biomacromolecules, 2003. Please note that modification of the recommended protocol may alter final gel properties. Therefore it is the responsibility of the end user to ensure reproducibility of the protocol.

Can I add more cell suspension and less buffer when resuspending QGel Matrix powder?

The standard protocol suggests to add 400 uL QGel Buffer A and 100 uL cell suspension for a total of 500 uL resuspension volume. The cell suspension volume can be lowered but QGel Buffer A volume has to be increased accordingly in order to keep the 500 uL total resuspension volume. Similarly, if the volume of the cell suspension is increased the QGel Buffer A volume has to be decreased accordingly (to keep the 500 uL total resuspension volume). Please take note that this may alter the gelation conditions of the QGel Matrix. Consequently, the final gel characteristics may be different than those presented by QGel.

How can I prevent the gels from sticking to the Disc Caster when harvesting?

It is essential to apply PBS (Phosphate-Buffered Saline) around the gel discs before opening the QGel Disc Caster. Then, make sure the PBS has wet the entire side surfaces of each gel disc. Use a spatula to spread the PBS all around gels. To avoid gel sticking to the caster surfaces, first detach slightly the disc edges with the spatula to allow the PBS to flow through upper interfaces. Before lifting the upper (transparent) part of the caster, make sure the discs are fully detached from this upper part. Once this is done, use the spatula as a lever to lift up slightly the upper plate of the Disc Caster. View the related video found in the Protocol Tab on this Product Descritpion page. For more information about how to make gel discs from the QGel Matrix powder, visit: https://www.qgelbio.com/files/protocols/Casting3D_Discs.pdf However, if making gel discs really doesn’t suit your experiment, you can also cast gel drops (i.e. without using the transparent cover) that are much easier to remove from the lower (white) part of the gel caster. Wet the gel drops with the PBS, harvest them with a spatula and put them in cell culture media.

Can I directly pipette QGel solution in multiwell plates (and thereby avoid making gel discs or drops)?

If you are looking to encapsulate cells in assay plates for high-throughput purposes, consider the QGel Assay Kits made available for each QGel formulation. QGel does not recommend casting QGel Matrix solution directly in multiwell plates, but it does recommend making discs or drops that freely swell and float in medium. QGel Matrices may swell slightly after reaction and if casted in a well, gel swelling may be impeded, resulting in gels with different material characteritics from those provided by QGel. The advantage of making gel discs or drops is that they can be easily transfered to new type of support and handled for different analysis (e.g. confocal, viability test, histology). Moreover, gas and nutrient diffusion is improved when discs can freely move in the well. It is very easy to make 3D gel discs or drops from QGel Matrix. You will find details instructions at: https://www.qgelbio.com/files/protocols/Casting3D_Discs.pdf

Are QGel Matrices injectable?

QGel matrices carrying cells can be transplanted to animal tissues to induce and study specific in vivo responses. Please refer to our line of in vivo matrices and the accompanying protocols or contact us for more information.

How can I assess cell viability and proliferation in QGel Matrices?

Conventional assays, such as live/dead stainings (e.g. Viability/Cytotoxicity Kit (Invitrogen)) and other qualitative/quantitative tests that involve spectroscopic measurements (e.g. Alamar Blue (Invitrogen)), can be used to assess cell viability and proliferation in QGel Matrices. Please visit the Protocol tab on the product description page.

Is it possible to recover cells from QGel Matrices after encapsulation?

For sub-culture, cells can be recovered from QGel Matrices by proteolytic digestion of the gels with trypsin. Conditions have to be optimized for each cell type and experimental setting. We can provide protocols for this. Please visit the Protocol Tab on this Product Description page.

Is it possible to extract DNA/RNA from cells encapsulated in QGel Matrices?

For DNA extraction, you can use a conventional kit prepared for measuring DNA (e.g. CyQUANT® or PicoGreen®) from Invitrogen. To extract/lyse the cells from the gels after cell culture, freeze the gel discs, and subsequently use proteases (e.g. Proteinase K) to dissolve the gels and lyse the cells. For RNA extraction, collagenase digestion, or alternatively mechanical destruction, of gels in appropriate RNA extraction buffer can be used to solubilize the gel for this purpose. We can provide protocols for this. Please visit the Protocol Tab on this Product Description page.

Can I do an immunostaining on cells encapsulated within QGel Matrices?

QGel Matrices are fully transparent and encapsulated cells can be observed clearly after weeks, even months, of culture depending on the density of the proliferating cells. Both conventional light microscope and confocal laser microscope (fluorescence staining) can be used. QGel Matrices allow for conventional immunofluorescence labelling of cells in gels and high-quality imaging using confocal laser microscopy. Histology can also be performed on QGel Matrices. Frozen section methods of gels are preferable (Adelöw, Biomaterials, 2008). Click here for the protocol for making frozen sections for histology We can provide protocols for this. Please visit the Protocol Tab on this Product Description page.

How is QGel Matrix in 2R vial format supplied?

QGel Matrices delivered in vial format as a lyophilized powder contain all pre-mixed components. One vial allows you to cast 500 ?l gel. Upon arrival, it must be stored at -20°C. In order to resuspend the powder into gel formation, QGel Buffer A will be included with each order. One buffer vial contain 4mL buffer each (0.3M HEPES, pH 7.9 ± 0.1) and can be used to resuspend as many as eight QGel Matrix powder vial. It must be stored at 4°C. The QGel Matrix powder and Buffer A are shipped in different boxes to facilitate storage and can be easily piled up in fridge/freezer. For more information about how to make gel discs from the QGel Matrix powder, click here: https://www.qgelbio.com/files/protocols/Casting3D_Discs.pdf

How is QGel Matrix in Assay Kit format supplied?

QGel Matrices are delivered as a lyophilized powder and contain all pre-mixed components in our 96-well Assay Kits. Each mini-vial in the Assay Kit yields 100 ?l gel once the gel has been resuspended and prior to dispension to assay plates. The respuspended gel and cell suspension is then dispensed into ca. 16 assay plate wells (based on 384-well assay plates). The Assay Kit must be stored at -20°C. In order to resuspend the powder into the gel matrix, QGel Buffer is added the the lyophilized formulation and then dispensed into separate assay plates. Please view the video in the Protocol Tab of this page. We will provide you with complete protocols with your order.

What is the storage temperature for the different QGel products?

QGel Matrix powder in 2R vial and Assay Kit format must be stored at -20°C. QGel Buffer A must be stored at 4°C.

How long can I store QGel products before they expire?

QGel Matrices have a shelf life of 5 years. They must be stored at -20°C. QGel Buffer A can be stored up to one year at 4°C.

How do I get additional information on QGel Matrix products (e.g. certificate of analysis, MSDS sheet, ...)?

Please contact us if you require a Certificate of analysis and MSDS for your QGel formulation. Certificate of analysis gives batch specific information (Lot number of QGel Matrix products is written on the label of each QGel formulation).