General
How Can I purchase?
Please visit our SHOP tab to purchase through our website, or see "Distributors" to order.
We accept PO numbers. Please email [email protected] to place an order.
Can't find us in your Vendor System? Email [email protected] and we will setup the account.
We accept PO numbers. Please email [email protected] to place an order.
Can't find us in your Vendor System? Email [email protected] and we will setup the account.
Do you have a refund policy?
We will replace any unused items that are not functioning or damaged upon delivery.
If you are unsatisfied with the product we will accept a return with a restocking fee.
If you are unsatisfied with the product we will accept a return with a restocking fee.
SeedEZ
What types of cells can be grown in SeedEZ?
We haven't found a cell type that has not been compatible with SeedEZ. From cell lines, to primary cells, to iPSC-derived cells, everything has thrived in our scaffolds. Please see our complete list of cells tested here: Cell Types. If your cell type is not on the list, please contact us and we would be happy to help optimize protocols for you!
Primary cells:
iPSC and iPSC-Derived Cells:
Immortalized Cells Lines:
Primary cells:
- Human Hepatocytes
- Human Astrocytes
- Human Brain Microvascular Endothelial Cells
- Human Brain Pericytes
- Human T Cells
- Human Lung Tumor Cells (Non-Small Cell Lung Carcinoma)
- Human Fibroblasts
- Human Neutrophils
- Rat Neurons (From Embryonic Day 18)
- Rat Astrocytes and Microglia (From Postnatal Day 0)
- Rat Neurospheres
iPSC and iPSC-Derived Cells:
- Human Hepatocytes
- Human Neurons
- Human Neural Progenitor Cells (NPCs)
- Human Astrocytes
- Human iPSCs
Immortalized Cells Lines:
- HEK293 (Human Kidney)
- BT-474 (Human Breast Cancer)
- SUM159 (Human Breast Cancer)
- HB2 (Human Breast Cancer)
- SKBR3 (Human Breast Cancer-Metastatic)
- Hep G2 (Human Hepatocytes)
- HepaRG (Human Hepatocytes+Biliary Cells, Terminally Differentiated)
- H1299 (Human Lung Cancer) HUVEC (Human Umbilical Vascular Endothelial Cells)
- HMC3 (Human Microglia)
- NIH-3T3 (Mouse Fibroblasts)
- MC3T3-E1 (Mouse Osteoblast Precursor Cells)
- Jurkat (Human T Cells)
- C2C12 (Mouse Myoblasts)
- Cal 27 (Human Head & Neck Squamous Cell Carcinoma)
- HUVEC (Human Umbilical Vascular Endothelial Cells)
Can I extract protein or RNA from cells in SeedEZ?
Yes. You may use your normal protocols for protein or RNA recovery directly in the SeedEZ scaffold. Remove any medium from the well. Gently perform rinsing steps to remove any residual medium from the scaffold. Add the minimum volume of lysis/extraction buffer necessary to cover the scaffold (note that residual liquid in the scaffold may require the use of more concentrated lysis/extraction buffer or a longer incubation time). If a hydrogel was used, it may be necessary to chemically or enzymatically dissolve it prior to exposure to lysis/extraction buffer.
Can I get cells out of SeedEZ?
Yes. Use your normal reagents for passaging cells directly on the SeedEZ scaffold. Recovery efficiency is variable and dependent on cell type and may require optimization.
Example:
Example:
- Remove any medium from the well. Gently rinse with PBS (without calcium and magnesium)
- Add trypsin to the well with scaffold and incubate as you normally would for passaging (e.g. 37 degrees Celsius for 5 minutes)
- Transfer scaffold to a 15 mL or 50 mL conical tube (depending on scaffold size) containing 10 mL (for 15 mL tube) or 20 mL (for 50 mL tube) PBS (calcium and magnesium-free)
- Centrifuge a 1000 rcf for 5-10 minutes. The SeedEZ scaffold will get caught in the conical portion of the tube; cells will be pelleted below it
- Remove the scaffold with sterile tweezers
- Aspirate supernatant
- Resuspend pellet with desired buffer/medium
Why is it recommended to pre-wet SeedEZ?
This step is optional, but recommended. Pre-wetting the scaffold ensures a
more homogenous distribution of cells within the scaffold. The random
orientation of the glass fibers can result in slightly inconsistent densities
throughout the scaffold if the cells in medium are added directly to the scaffold.
Pre-wetting makes the scaffold even more hydrophilic, and the cells in medium
will be able to distribute evenly.
more homogenous distribution of cells within the scaffold. The random
orientation of the glass fibers can result in slightly inconsistent densities
throughout the scaffold if the cells in medium are added directly to the scaffold.
Pre-wetting makes the scaffold even more hydrophilic, and the cells in medium
will be able to distribute evenly.
How many cells can i plate?
We recommend seeding no fewer than 5,000 cells, below are our recommendations for seeding volume and density:
SeedEZ Size |
Seeding Volume (μL) |
Seeding Vol. Hydrogel (μL) |
Cell Density |
Medium Volume |
6 Well |
360 |
360-450 |
9.0E5 - 9.0E6 |
2.00 mL |
12 Well |
140 |
14-175 |
3.5E5 - 3.5E6 |
1.00 mL |
24 Well |
80 |
80-100 |
2.0E5 - 2.0E6 |
500 μL |
48 Well |
40 |
40-50 |
1.0E5 -1.0E6 |
250-400 μL |
96 Well |
10 |
10-15 |
3.0E4 - 1.5E5 |
150-200 μL |
Can i buy the scaffolds in sizes other than the ones listed?
We offer SeedEZ scaffolds for most multi-well plate formats (6, 12, 24, 48, and 96 wells). We also offer a 5 x 10 cm sheet from which you can cut your own using sterile scissors. We can create custom shapes and sizes upon request. Let us know!
Can I transfect cells in SeedEZ?
Yes. You can transfect cells in SeedEZ. Note that you many need to increase the amount of plasmid delivered to correspond to the number of cells seeded in the scaffold.
Can I use SeedEZ as an implant?
No. SeedEZ scaffolds are for in vitro research use only.
How do I image cells in SeedEZ?
Although SeedEZ scaffolds are made of transparent glass microfibers, they appear phase-dark with phase-contrast microscopy. We recommend fluorescence microscopy for imaging cells in the SeedEZ scaffold. Cells expressing fluorescent markers or stained with live/dead stains such as Calcein AM and propidium iodide can easily be imaged using epifluorescence or confocal microscopy. Cells may also be fixed in situ and processed for immunocytochemistry using fluorescent secondary antibodies.
PerfusionPal
What is included in the Starter System?
The Starter System includes a dual syringe pump, two bottles of Blood Substitute (100 mL each), and two complete PerfusionPal systems (each containing: plate, seating insert, scaffolds, tubing, and a luer cap for non-perfused applications).
Is there any cross-talk between the wells?
No. The blood substitute does not dissolve anything from the cell culture medium. Each well of the PerfusionPal is completely independent.
Can I reuse the perfusionpal plates?
No. PerfusionPal is a one-time use product. However, the blood substitute is completely reusable. We do offer discounts on 4-packs of our PerfusionPal plates. For high-volume orders, email [email protected] for discount pricing.
How do i reuse the blood substitute?
Blood Substitute can be reused indefinitely.
After use, remove the multi-well insert from the PerfusionPal and allow to drain on absorbent bench paper. Remove any medium or buffer from the surface of the Blood Substitute. Collect the Blood Substitute from the tray, tubing, and syringe. Sterile filter with a 0.2 µm filter prior to use in a subsequent experiment.
Prior to biohazard disposal, allow residual blood substitute to evaporate from the tray.
Do not discard the Blood Substitute down the drain or dispose of with waste that will be incinerated.
After use, remove the multi-well insert from the PerfusionPal and allow to drain on absorbent bench paper. Remove any medium or buffer from the surface of the Blood Substitute. Collect the Blood Substitute from the tray, tubing, and syringe. Sterile filter with a 0.2 µm filter prior to use in a subsequent experiment.
Prior to biohazard disposal, allow residual blood substitute to evaporate from the tray.
Do not discard the Blood Substitute down the drain or dispose of with waste that will be incinerated.
Can I use a different pump with PerfusionPal?
Yes. Other syringe pumps that are capable of both infusion and withdrawal may be used with PerfusionPal. However, we cannot offer technical support for pumps other than the one included with the Starter System.
How do I change the medium?
We recommend exchanging half of the volume of medium in each well every 2-3 days when the perfusion system is fully infused (syringe plunger is pushed in to the 5 mL mark). Remove half of the medium volume with a micropipette and replace with the same volume of fresh medium.
Since some of the medium will be below the scaffold and inaccessible, it is not feasible to remove it all. Additionally, our research has shown that 3D cultures grow better when they are not exposed to a complete change in medium. Removal of secreted signaling molecules and growth factors can have a detrimental effect on the culture.
For end-point assays that require complete removal of medium, we recommend removing the scaffolds from the PerfusionPal and transferring them to fresh medium/buffer in a multi-well plate.
Since some of the medium will be below the scaffold and inaccessible, it is not feasible to remove it all. Additionally, our research has shown that 3D cultures grow better when they are not exposed to a complete change in medium. Removal of secreted signaling molecules and growth factors can have a detrimental effect on the culture.
For end-point assays that require complete removal of medium, we recommend removing the scaffolds from the PerfusionPal and transferring them to fresh medium/buffer in a multi-well plate.
Why are the perfusion rates so low?
PerfusionPal is designed to model the interstitial fluid flow rates in the human body. The recommended settings for the syringe pump correspond to 16 culture volume exchanges per day. We have found that slower flow rates result in enhanced cell viability and function. Should you require higher flow-rates for your studies, the PerfusionPal system can easily accommodate rates that allow up to 600 culture volume exchanges per day.
Does the pump go in the incubator? / Where does the pump go?
No. the syringe pump remains outside of the incubator. The PerfusionPal plate is placed in the incubator with the tubing placed between the door and the gasket. The tubing is equipped with a steel sheath that should be placed where the door meets the gasket to ensure the tubing is not crushed. The syringe pump should be positioned on a stand or stool such that it is at or below the level of the PerfusionPal plate.
Can I grow tissue slices in PerfusionPal?
We are currently developing protocols for maintenance of tissue slices in PerfusionPal.
Can I image cells in PerfusionPal?
We recommend removing scaffolds from the PerfusionPal and transferring them to a multi-well plate for fluorescent labeling and imaging.