521-To-Go™

521-To-Go™ - Laminin-521 precoated 24-well plates  


Easy stem cell culture just became easier with 521-To-Go plates. 521-To-Go allows you to concentrate on your science, not on technical issues like coating plates. 

Laminin-521 (LN-521) is the natural laminin for pluripotent stem cells and therefore reliably facilitates self-renewal of human ES and iPS cells in a chemically defined, feeder-free and xeno-free stem cell culture system. LN-521 enables efficient single-cell passaging of genetically stable and pluripotent stem cells without need of any apoptosis inhibitors for superior quality of your cells and studies. Due to the simplicity of the passaging procedure the experimental variance is reduced and the process can be automated for high-throughput stem cell culture and scientific results.

After birth, α5-laminins represent the most common laminins in the body and LN-521 supports many diverse tissue cell types, such as cells from pancreas, vascular, nervous and muscular systems.

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Amount
Size
Price
Oty
24 well 1 plate €67
24 well 5 plates €298
24 well 20 plates €1055
For larger quantities contact us

Estimated shipping time
1-5 working days

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Human embryonic stem cells have been considered difficult to culture by most people since they were first derived in 1998. First cells were cultured on top of other cells, which was extremely labor intensive, then on top of different matrices where cells behaved sub-optimally.

Human recombinant LN-521 revolutionized stem cell culture. Suddenly culturing pluripotent stem cells was easy, efficient and reliable.

LN-521 recreates the natural environment of the blastocyst and therefore cells stay pluripotent without karyotypic changes for more than 130 passages.

LN-521 enables clonal derivation of hESC lines, clonal survival of hESCs and efficient expansion of single-cell plated hPSCs under completely chemically defined, xeno-free conditions (Rodin, 2014).

Long-term expansion of hPSCs with LN-521 can be done in a chemically defined and xeno-free system with high cell expansion rate, monolayer growth, maintained pluripotency and most importantly with normal karyotypes.

 

LAMININ-521 CELL CULTURE APPLICATION NOTE

Clonal survival enabled by LN-521 of hESCs and iPSCs will greatly facilitate the study of genomic alterations and manipulations in pluripotent cells. Furthermore, renders more efficient reprogramming of specific tissue cells to iPSCs (Lu HF, 2014).

Derivation of new, genetically stable hESC lines with LN-521 is performed in a chemically defined, xeno- and feeder free manner, which can therefore be used for the development of cell lines for cell therapy. The method can even be employed on single blastomeres from IVF embryo biopsies, thus will not destroy the embryo.

LN-521 stem cell matrix will help your research and the development of cell therapy for humans. However, the coating of cell culture plates with LN-521 still takes time. 521-To-Go™ solves this problem. Now you can just open the bag, take out your cell culture plate and start culturing your stem cells directly. This allows you to concentrate on your scientific questions instead of technical problems!

 

 

 

Authentic human pluripotent stem cell culture

Human stem cell culture

Easy culture of human ES and iPS cells in a 
biorelevant, defined and xeno-free environment 

Authentic clonal cell culture

Clonal culture of hPSCs

Efficient clonal culture and derivation of
human pluripotent stem cells 

SINGLE-CELL PASSAGE
WITH 521-To-Go™

1. Aspirate old medium from wells and wash the cells gently with 1xDPBS (Ca--/Mg--) once.

2. Add any enzyme (ie TrypLE™) or 1mM EDTA diluted in 1xDPBS and incubate at +37°C, 5% CO2 for 3-5 minutes (6-8 minutes with EDTA) to detach the cells.

3. Add same volume of enzyme inhibitor if appropriate or fresh medium. Pipette up and down 6-10 times to achieve single-cell suspension (if appropriate).

4. Collect the single-cell suspension in conical tube containing 1ml of fresh medium, centrifuge at 800 rpm for 4 minutes and discard supernatant.

5. Re-suspend the cell pellet in 1 ml of fresh medium.

6. Count the cell number. Seed cells with density of 50,000 cell/cm2 or with a split ratio of 1:10 to 1:30.

7. Swing the plates side-to-side to distribute cells evenly, then place them into an incubator (+37°C, 5% CO2).

8. Feed cells on daily basis until next passage. Add only a few drops of fresh medium after 24 hours and perform a complete medium change 48 hours after passaging.

 

Instruction for cell culture on 521-to-go plates

 

SIMPLE PROCEDURE WITH 521-To-Go™

1. Just open the bag under laminar flow hood.

2. Add culture medium to the wells and equilibrate at +37°C, 5% CO2 during passaging procedure until cells are ready to be seeded. The pre-coated well does not require washing befor use.

3. Seed your human PSCs at appropriate density and incubate at +37°C, 5% CO2.

IMPORTANT NOTES

  • The Laminin pre-coated plates are stable for 1 year when stored at +15°C to +25°C.

  • The pre-coated wells do not require washing before use. 

  • When using LN-521 no treatment with apoptosis inhibitors, such as ROCKi or blebbistatin, is needed. 

  • The protocol can easily be made totally defined and xeno-free with your choice of culture medium and enzyme.

  • Before start, all solutions used for cell passaging should be aliquoted in sufficient amounts and pre-warmed at +37°C, 5% CO2.

  • Cells are ready to be passaged when cell culture is ≥60% confluent. Optimal seeding densities will vary from one cell line to another and can be determined empirically for your system. With optimal media conditions and seeding density, most cell lines will reach confluence within 4-6 days and expand 10-25 fold.

  • For cell densities lower than 5,000 cells/cm2 or clonal assays, ROCKi should be added.

  • When moving your cells from another feeder-free matrix (e.g. Matrigel) we recommend you to start smaller well format (e.g. 96-well or 48-well format) and a higher seeding density (50,000-100,000 cells/cm2) for the first number of passages to let the cells adapt to the laminin matrix before increasing the well format and lowering the seeding density.

  • When moving your cells from feeders to LN-521, follow the protocol decribed here

 

 

 

Product name

521-To-Go™

 

Product use

Cell culture

  


 

Documents

MSDS

Storage

Room temperature (+15°C to +25°C)

 

Stability

One year from manufacturing date

 

Product application

Human embryonic and induced pluripotent stem cells

 

Protein name

Laminin-521 (LN-521)

 

Synonyms

Laminin-11

 

Expressed in vivo

Expressed and secreted by human pluripotent stem cells and can, in addition, also be found in the kidneys, neuromuscular junctions, lungs and placenta.