Biolaminin 521 MX

Amount	Size	Price	Oty
500 ug	1 x 5 ml	€258
For larger quantities contact us

Applications Supporting data Instructions Technical info Publications

Features and Specifications:

Research grade substrate
Animal component-free to the secondary level
Biologically relevant culture environment
Homogenous and genetically stable hPSC cultures
An easy and flexible culture system
Consistent and reliable performance
Rapid scale-up
Easily adaptable to automation platforms
Supports a weekend-free feeding regime
Efficient differentiation and enhanced cell maturation, polarization, and organization
Scientifically proven

A BIOLOGICALLY RELEVANT CULTURE ENVIRONMENT

Biolaminin 521 MX is a full-length, human, recombinant laminin 521 substrate, the only one of its kind on the market, providing an optimal environment for feeder-free culture of hPSCs. Laminin 521 is a key cell adhesion protein of the natural stem cell niche, expressed and secreted by hPSCs in the inner cell mass of the embryo. The Biolaminin 521 substrate recreates a biologically relevant hPSC milieu in vitro, promoting attachment, high survival, and robust long-term self-renewal of hPSCs. In addition, the more authentic culture environment primes the pluripotent stem cells for more efficient differentiation, enhanced cell maturation, polarization and organization of specialized cell types. The laminin 521 cell culture substrates have a positive effect on stabilizing and homogenizing pluripotent gene expression profiles between hES cell lines, providing a more controllable and robust culture system compared to the feeder- and many other feeder-free culture systems. Via the interaction with relevant cell surface receptors, laminins activate cell signaling cascades (e.g. PI3K/Akt pathway), giving consistent cellular responses and improved cell functionality, ultimately saving researchers both time and cost.

The MX521 stem cell substrate enables high expansion rate, monolayer homogenous growth, and maintained pluripotency. Cells are genetically stable without karyotypic abnormalities. hESC and iPSC grow faster on MX521 compared to cells cultured on feeders or on another feeder-free matrice, facilitating automation. Human pluripotent stem cells can routinely be cultured as single cells without the addition of ROCKi. MX521 also supports efficient clonal survival and expansion of hES and iPS cells, greatly facilitating the study of genomic alterations and manipulations in pluripotent cells. Furthermore, it renders more efficient reprogramming of specific tissue cells to iPSCs.

The Biolaminin 521 substrate is more flexible, easier to use and less laborious compared to feeder-based culture systems and is compliant with any commercially available serum-free medium. The cells grow in a homogeneous monolayer and are easy to monitor, with maintained pluripotency and genetic integrity. The robust and flexible culture environment allows operator-independent culture maintenance and reliable, standardized protocols, which can easily be adapted to automation platforms.

After birth, laminin 521 represents one of the most common laminins in the body and together with the other laminin isoforms, it supports efficient expansion and differentiation of many diverse tissue cell types.

Download cell culture applications poster

Human stem cell culture

Easy culturing of human ES and iPS cells in a
biorelevant and animal component-free environment

effective clonal culture

Clonal cell culture

Efficient clonal culture and derivation of
human pluripotent stem cells

COATING PLATES

See instructions for coating protocol, recommended coating volumes, and concentrations.

hPSC CULTURE ON MX521

See instructions for stem cell culture on Biolaminin 521, including:
- transfer
- passaging
- thawing
- cryopreservation of hPSCs on MX521.

Embryoid body formation

Protocol for embryoid body formation from hPSCs cultured on Biolaminin 521 substrates

IMPORTANT NOTES

"For Research Use Only"
Avoid long exposure of the protein to ambient temperatures
CT521 is animal-origin free to the secondary level according to the ISCT guidance document ˮISCT Animal-Free Origin Survey Results-Summary”.
The laminin stock solution is stable for 3 years when stored at -20°C to -80°C. Thawed, undiluted laminin stock is stable for at least 3 months when stored at +2°C to +8°C under aseptic conditions.
Repeated freeze-thawing should be avoided. If desired, the laminin stock can be dispensed into working aliquots and stored at -20°C to -80°C.
For your convenience, the coated plates can be kept for up to 4 weeks when stored aseptically at +2°C to +8°C
The protocols can easily be made totally defined and animal component-free with your choice of culture medium and dissociation reagent
It is important that the cells transferred to the MX521 matrix are of high quality
Some hPSC lines transferred to the MX521 matrix, might require an adaptation period before they can be cultured according to the single-cell passaging protocol
Once adapted to the MX521 matrix, hPSCs can routinely be cultured as single cells without ROCKi

Troubleshooting

Biolaminin plate coating

An uneven cell spread, it’s often a coating issue and could be caused by the following:

To low coating concentration used. Increase the coating concentration is high enough to support even cell growth.
Bad coating coverage/the plate has dried out. Ensure that the entire surface is covered by the laminin coating solution when preparing fresh plates. Also, do not let the plate dry out as this will inactivate the laminin coating. Too long time in the incubator or long storage without sealing could cause too much evaporation so that part of the plate dries out (often in the center).

Transfer from other substrates

We recommend to transfer the cells as single cells (or as small clumps) and always with the addition of ROCKi for the first few (3-5) passages. Once the cells are adapted to the MX521 matrix, the cells can be cultured as single cells without ROCKi. This may take up to 5 passages.
If the cells are hard to adapt, try increasing the coating concentration to 10 ug/ml and seed at a higher cell density 50,000 – 100,1000 cells/cm². Once the cells are adapted a lower coating and seeding concentration often can be used.
It is important that the cells transferred to the MX521 matrix are of high quality. MX521 will generally also support differentiated cells so carefully select only undifferentiated cell areas for transfer.
MX521 works well in combination with most commercial media brands. However, it is to be expected that cell morphology will look different depending on the medium used for culture.

hPSC splitting and seeding

We recommend single-cell passage or passage as small clumps.
Stem cells are sensitive and when using an enzyme, do not treat the cells too long as that will damage the cells. Cells attach tighter on laminin compared to other matrices and scraping or pipetting without first loose up cells can affect cell integrity and viability which could result in less attachment the next day. Less confluent cells need shorter treatment time whereas more confluent cells might need longer treatment time. The cells should detach easily without too much pipetting needed. Do not use too much mechanical force (extensive pipetting or scraping) as that will damage the cells. Increase the dissociation reagent incubation time rather than increasing the force. If the cells stick very hard to the MX521 surface, try to lower the coating concentration.
After seeding: most cells should have attached after 1 hour and the cells should be evenly distributed over the entire plate. If there is a lot of cell death after seeding, the cells have most likely been treated too harshly during splitting.
The day after seeding: the cell has migrated and should have formed small colonies and will continue to expand as a homogenous monolayer. Cells cultured on the MX521 matrix are ready to be passaged when cell culture is 60-99% confluent. Depending on the cell line, seeding density and on the medium used, cultures are usually passaged 3-6 days after seeding.

Product name

Biolaminin 521 MX

Product code

MX521

Declaration

For research use only

Documents

MSDS

CoA

Storage

-20°C to -80°C

Concentration

0.1mg/ml

Appearance

Clear, colorless, solution with a pH of 7.5 with
20mM HEPES, 0.25M NaCl, 10% glycerol

Shipping Condition

Dry Ice

Protein name

Laminin 521 (Laminin-11)

Classification

Animal component-free, human
recombinant protein

Product application

Culture of human embryonic and induced pluripotent stem cells (hESC and hiPSC), mesenchymal stem cells (MSC) and most anchorage-dependent progenitor cell types. Differentiation and maintenance of specialized cells, such as hepatocytes, cardiomyocytes, and neurons.

Clonal culturing of human embryonic stem cells on laminin-521/E-cadherin matrix in defined and xeno-free environment

Rodin S., Antonsson L., Niaudet C., Simonson O.E., Salmela E., Hansson E.M., Domogatskaya A., Xiao Z., Damdimopoulou P., Sheikhi M., Inzunza J., Nilsson A.S., Baker D., Kuiper R., Sun Y., Blennow E., Nordenskjöld M., Grinnemo K.H., Kere J., Betsholtz C., Hovatta O., Tryggvason K.
Nature Communications, 2014

Monolayer culturing and cloning of human pluripotent stem cells on laminin-521 based matrices under xeno-free and chemically defined conditions

Rodin S., Antonsson L., Hovatta O., Tryggvason K.
Nature Protocols, 2014

a-5 Laminin Synthesized by Human Pluripotent Stem Cells Promotes Self-Renewal

Laperle A., Hsiao C., Lampe M., Mortier J., Saha K., Palecek S.P., and Masters K.S.
Stem Cell Reports, 2015

Laminin 521 stabilizes the pluripotency expression pattern of human embryonic stem cells initially derived on feeder cells

Albalushi H., Kurek M., Karlsson L., Landreh L., Rós Kjartansdóttir K., Söder O., Hovatta O., Stukenborg J-B.
Stem Cell International, 2017

Niche-derived laminin-511 promotes midbrain dopaminergic neuron survival and differentiation through YAP

Zhang D., Yang S., Toledo E.M., Gyllborg D., Saltó C., Villaescusa J.C., Arenas E.
Sci Signal. 2017