Is FN1 motifs a two-dimensional surface?
Yes, FN1 motifs is a two-dimensional surface.
What peptide sequences are used to mimic the ECM?
The FN1 motifs surface is made up of synthetic fibronectin-derived motifs including RGD peptide sequence.
Is FN1 motifs also available for manual coating?
A FN1 motifs substrate for manual coating is currently not available.
How is FN1 motifs manufactured?
The FN1 motifs consumables are developed and manufactured according to ISO 9001. Production of all Eppendorf Cell Culture Consumables takes place in a controlled class 5 clean room environment according to ISO 14644-1 and EG-GMP Class A. They are manufactured using aseptic processing per ISO 13408-1 “Aseptic processing of healthcare products”. Each step was validated to ensure the FN1 motifs consumables meet the Sterility Assurance Level (SAL) 10-3. The consistent reliable quality of FN1 motifs consumables is ensured by monitored process controls and checks from the raw material to the final packed product. The growth surface is ensured to fulfill all requirements concerning homogeneous coating and to be animal- and xeno-free.
Does FN1 motifs have autofluorescence?
No, FN1 motifs cultureware does not present autofluorescence.
Do I have to prepare the FN1 motifs surface before use?
No, the FN1 motifs surface is ready-to-use and does not require any pre-equilibration or washing step before use. Keep the product sealed in original packaging and open it just before use.
I have already opened the package. How soon do I have to start working?
The package should be opened just prior to use.
Why is the FN1 motifs surface individually packed in aluminum foil?
The aluminum foil protects the FN1 motifs surface from the external environment (e.g. UV light) and ensures its stability and sterility throughout the shelf-life of 3 years at room temperature based on unopened product.
The FN1 motifs consumable is exposed to the laboratory environment without the aluminum-foil based packaging. Can I still use the surface?
We do not recommend using the product after vessels are exposed to the laboratory environment without the aluminum-foil based packaging. Stability and sterility cannot be guaranteed anymore.
Can I use any unused wells of a FN1 motifs plate?
Stability and sterility of unused wells cannot be guaranteed as it might be impaired during handling or incubation. Therefore, we recommend using a new sterile plate for the further experiment.
Should FN1 motifs be protected from UV light?
The aluminum-foil based packaging protects the FN1 motifs consumable from UV light. The package should be opened just prior use. The FN1 motifs surface should not be exposed to UV radiation, e.g. in the biosafety cabinet.
How can I store FN1 motifs cultureware?
The FN1 motifs cultureware should be stored in the original aluminum-foil based packaging in a dry room at room temperature at 15°C to 30°C. Its shelf-life of 3 years at room temperature is based on unopened product.
Can I store FN1 motifs consumables at 4°C?
FN1 motifs consumables in the aluminum-foil based packaging can be stored at 4°C, but have to be adjusted to room temperature before use.
What cell types can be cultured on FN1 motifs?
The FN1 motifs surface is suitable for culture of eukaryotic cells, especially pluripotent and multipotent stem cells, as well as for expansion and differentiation even in restrictive (e.g. serum-free) cell culture conditions. Please refer to the user manual
for a list of cell types already expanded on FN1 motifs.
Does FN1 motifs support a xeno-free culture of stem cells?
The FN1 motifs surface is fully xeno-free and free of any animal or human components. FN 1 motifs supports the culture of stem cells in commercially available xeno-free media in combination with xeno-free cell detachment solution. Please refer to the user manual
for a list of media and cell detachment solutions already used for xeno-free cell expansion on FN1 motifs.
What cell detachment solutions work with FN1 motifs?
FN1 motifs supports cell detachment with various reagents, e.g. several TrypLE™ reagents, Accutase®
, Versene, Dispase and Trypsin/EDTA. Please refer to the user manual
for a complete list of dissociation reagents already used for cell detachment on FN1 motifs. We recommend using the manufacturer protocols as provided.
Can I use the FN1 motifs surface for cell-based assay and stainings?
Yes, the FN1 motifs surface is compatible with cell-based assays and staining procedures.
Does FN1 motifs support the long-term expansion of stem cells?
Yes, FN1 motifs supports the long-term expansion of hiPSCs (at least 25 passages) and hMSCs (at least 10 passages). Please refer to our Application Notes 389 and 390 for further details on the long-term expansion of hiPSCs and hMSCs on the FN1 motifs surface, respectively.
Can I thaw hMSCs directly on FN1 motifs surface?
hMSCs can be directly thawed on FN1 motifs.
Can I thaw hiPSCs directly on FN1 motifs surface?
You can directly thaw hiPSCs on FN1 motifs if cells were originally frozen from the FN1 motifs surface. hiPSCs can be thawed either in clumps or single cells. If the cryopreserved cell stock was prepared from hiPSCs cultured in a different medium-surface culture system, we recommend thawing of cells in the original culture system and to initiate cell transition on FN1 motifs after full recovery from thawing. Please refer to the user manual
for detailed tips of hiPSC transition on FN1 motifs surface.
Do I need to add an apoptosis inhibitor for hiPSC seeding on FN1 motifs?
Supplement your media with ROCK inhibitor for 24 h post-seeding during the surface transition period. After adaptation to the new surface, the use of ROCK inhibitor is not required for long-term expansion using clump passaging. For single-cell passaging, the use of ROCK inhibitor for 24 h post-seeding is recommended during the entire expansion process.
What is the recommended hiPSC seeding density and split ratio on FN1 motifs?
Chose a higher initial hiPSC density for thawing and surface transition than usually used during your routine culture (split ratios of 1:4 to 1:6, corresponding to 1 to 1.5x105 cells per cm2). Decrease it progressively until complete cell adaptation on FN1 motifs is reached. After adaptation typical split ratios used for hiPSC expansion on FN1 motifs are 1:8 - 1:12 (corresponding to 5 to 7.104 cells per cm2). However, as optimal cell densities for seeding can vary based on cell lines and media used, seeding densities of hiPSCs can be also optimized by the user. We recommend using different split ratios in parallel to allow optimal cell population selection for next passage.
What is the recommended passaging rate for hiPSC expansion?
Depending on the used seeding densities, hiPSCs are typically passaged every 3 to 4 days on FN1 motifs when cell population reaches 70 to 80% confluency. Passaging should also be initiated if colonies become too dense or if enhanced spontaneous differentiation is observed. Avoid to passage cells too early or too frequently. Optimal passaging frequency should be optimized by the user based on hiPSC line and culture media.
What passaging techniques of hiPSCs can be used with FN1 motifs?
FN1 motifs is compatible with either clump or single-cell passaging methods for routine hiPSC expansion. Mechanical passaging using sterile cell scraper or pipette tips is also feasible. Please refer to the user manual
for a list of detachment solutions already used for cell detachment on FN1 motifs. For surface transition, clump passaging using Versene, EDTA or Gentle solutions is recommended. Cell passaging method could be changed once cell culture is stable on the new surface.
How do hMSCs look like when cultured on FN1 motifs?
hMSCs cultured in traditional serum-containing medium display their characteristic fibroblast-like morphology on FN1 motifs. The FN1 motifs surface also efficiently supports hMSC attachment and growth in various serum-free culture systems, showing a more elongated spindle shaped cell morphology typical for serum-free culture conditions, more compact monolayers and higher cell densities.
How do hiPSCs look like when cultured on FN1 motifs?
The morphology of hiPSCs expanded on FN1 motifs corresponds to the hiPSC morphology expected on a feeder-free culture system, characterized by compact cells with a high nuclear-to-cytoplasm ratio and a prominent nucleolus, forming flat, tightly packed, shiny colonies with well-defined boarders.
Do stem cells previously cultured on a different surface undergo an adaptation period after they are transferred on FN1 motifs?
Stem cells can be directly seeded on FN1 motifs. The original stem cell population should be stable and robust prior transition. hiPSCs might need to adapt to the FN1 motifs surface during the first passages. Please refer to the user manual
for detailed tips of hiPSC transition on FN1 motifs surface.
I would like to transfer hiPSCs from a feeder layer-dependent surface on FN1 motifs. How should I proceed?
For surface transition from feeder-dependent surfaces, dislodge colonies using the usual cell dissociation method. Prefer homogeneous colony fragment size avoiding too small fragments. Medium supplementation with ROCK inhibitors (e.g. Y27632 at 10 µM) for 24 h post-seeding is recommended during surface transition period. After adaptation to the new surface, the use of ROCK inhibitors is not required for long-term expansion using clump passaging. For single-cell passaging, the use of ROCK inhibitors for 24 h post-seeding is recommended during the entire expansion process.
Where do I find the order number and the lot number?
You can find the order number as well as the lot number on the label of the aluminum-based packaging as well as on the outer box packaging.
Are your products certified sterile, free from Endotoxins and DNA, DNase and RNase?
Yes, all Eppendorf Cell Culture Consumables are certified non-cytotoxic (absence of pyrogens) and free of human and bacterial DNA, DNase and RNase. For detailed information please refer to the respective certificate of quality and the lot-specific certificates.