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.
Are your products free of BSE/TSE?
Eppendorf only works with granulate suppliers who guarantee that their used animal components come exclusively from countries without BSE occurrences. Risky materials are not used. The granulate production includes process steps with conditions above 235°C/ 3,000EkPa for several hours that destroy any virus, bacteria or substance that cause immunological diseases (TSE, BSE, CJD). For detailed information please refer to the respective certificate of quality and the lot-specific certificates.
What are the recommended working volumes for the different formats?
For information regarding working volume, maximum volume, and growth area of the different formats please refer to the Eppendorf CCCadvanced FN1 motifs user manual chapter 4/technical data.
For which applications may I use Eppendorf Conical Tubes® 25 mL?
In general, all applications where standard conical tubes 15mL and 50 mL are routinely used. The Eppendorf Conical Tubes® 25 mL close a gap between 15 mL and 50 mL conical tubes and additionally offer in this format a unique snap cap – the so called SnapTec™ cap - which markedly improves handling. This makes the Eppendorf Conical Tubes® 25 mL ideally suited for all cell/tissue culture applications including splitting cells, media change, transfections, etc. Also assay preparation and biomolecule purification protocols (gDNA, plasmid DNA, RNA, proteins), which require organic solvent extractions and multiple centrifugation steps may be successfully performed. They are also ideally suited for general reagent / sample handling and space-saving storage.
Is it necessary to buy new equipment to enable the use of Eppendorf Conical Tubes 25 mL within existing lab periphery?
No. The bottom shape / diameter of the Eppendorf Conical Tubes 25 mL is virtually the same with standard conical 50 mL tubes. With the respective height adapter (provided as Eppendorf accessory), the Eppendorf Conical Tubes 25 mL will be compatible with a wide range of standard laboratory equipment: centrifuges, mixers, etc., additionally tube racks, storage boxes and single-tube stands (skirts) are available for lab bench work, weighing and storage. For detailed accessories description please refer to www.Eppendorf.com .
Is it possible to store samples in 25 mL Eppendorf Conical Tubes in the freezer?
Yes, the tubes can be stored down to -86 °C.
How resistant are the Eppendorf Conical Tubes® 25 mL to chemicals?
The tubes are made of high quality, transparent polypropylene and resistant to a majority of commonly used chemical substances in the laboratory. An overview is provided in Application Note No. 56. In case a substance/chemical is not listed there, please contact our local contact.
Which purity grades and LoBind / Amber variants are available for Eppendorf Conical Tubes 25 mL?
The Eppendorf Conical Tubes 25 mL are available in the purity grades: Eppendorf Quality, PCR clean and Sterile/Pyrogen-free. For general and lot-specific certificates please refer to Eppendorf website eppendorf.com/certificates. The DNA and Protein LoBind as well as Amber variants of Eppendorf Conical Tubes® 25 mL will be available within short term as well.
Correct opening and closing of the snap Lid (SnapTec®).
The snap cap of the Eppendorf Conical Tubes® 25 mL is much larger than in other tube formats and may require some short learning of correct closing/opening at the beginning. Opening: press the safety tab from below and steady press upwards. In case of pressure buildup (organic solvents, higher temp incubations) make sure you slightly hold the lid with your thumb and don’t let the lid to pop out rapidly. Closing: press down the lid in the front area and make sure the safety tab clicks and is locked at right angle to the tube. Improperly locked tab may cause leaking.
How do Cell Culture Consumables enable gas exchange?
In Cell Culture Consumables gas exchange occurs via the vented lid design for plates or dishes or the integrated filter in filter cap flask. The Eppendorf plug-seal cap is equipped with a defined arrested and venting position that prevents unintended full closing.
How can I calculate the correct rotor speed (RPM) to use for pelleting my cells by centrifugation?
Most common protocols refer to 300 x g for 5 min to pellet eukaryotic cells. For information related to Eppendorf centrifuges and rotors, please refer to: Eppendorf centrifuges
Can I store my cell culture products in the biosafety cabinet during UV-Sterilization?
Eppendorf Cell Culture products are not resistant to longer UV-radiation with an intensity that is usually used for UV-sterilization of biosafety cabinets.
What is the temperature range the Eppendorf Cell Culture Consumables can be used?
Eppendorf Cell Culture Consumables are stable in the range of -86°C to 60°C.
Do the Eppendorf Cell Culture Consumables promote the attachment and proliferation of adherent cells?
We offer tissue culture (TC) treated or non-treated cell culture consumables. The TC treated products promote adhesion of adherent cells on the surface. The TC treatment procedure generates charged groups on the usually hydrophobic polystyrene surface. This facilitates cell attachment and spreading of most anchorage dependent cells.