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The Biolaminin 521 stem cell matrix

In the developing embryo, laminins are among the first extracellular proteins to be expressed. 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. Laminin 521 is also produced endogenously by pluripotent stem cells cultured in vitro and is a critical autocrine and paracrine factor that regulates their survival and self-renewal.

The Biolaminin 521 cell culture substrates successfully recreate the biologically relevant hPSC milieu in vitro. Via integrin binding, Biolaminin 521 induces the PI3K/Akt signaling pathway, promoting high survival and robust long-term self-renewal of human embryonic stem cells (hESC) and induced pluripotent stem cells (iPSC). The cells grow in a homogeneous monolayer with maintained pluripotency and genetic integrity and there is no need for manual removal of differentiated cell areas. With the defined and animal origin-free Biolaminin 521, the cell culture system culture of hPSC becomes easy, reliable, and standardized.   

KEY BENEFITS OF BIOLAMININ 521 FOR HUMAN PSC CULTURE

• Defined and animal origin-free cell culture matrix - for controlled cell culture and clinical compliance

• No lot-to-lot variability - more reliable and standardized experiments

• Homogenous hPSC cultures that are pluripotency and genetically stabile - no need to remove differentiated cell areas

• High clonal survival - optimal substrate for derivation and gene editing 

• Triggers authentic cell signaling pathways which generate hPSCs cultures with a more uniform gene expression profile

• High hPSCs expansion rate and controlled single-cell passaging - facilitates automation and high-throughput processes

• The authentic milieu that primes the hPSCs which results in more efficient differentiation and enhanced cell maturation, polarization, and organization

• Flexible culture system - easy to control and adaptable to any protocol

• Support weekend feeding - no more weekend work!

Download the application note for hPSC culture on the Biolaminin 521 LN (LN521) stem cell matrix.

Download the application note for the cell therapy grade Biolaminin 521 CTG (CT521).

FASTER EXPANSION OF hPSCs ON LN521

hESC and iPSC grow faster on Biolaminin 521 LN (LN521) compared to other feeder-free matrices, such as Vitronectin and EHS extracts (e.g. Matrigel), facilitating automation. The specific binding of LN521 to the α6β1 integrin enables high hPSC survival and self-renewal. PSCs on LN521 grow as a homogenous monolayer without any abnormal genetic aberrations. Once adapted to the LN521 matrix, hPSCs can routinely be cultured as single cells without the addition of apoptosis inhibitors, such as ROCKi. LN521 also supports high clonal survival and is a good substrate for derivation and gene editing.

NO SPONTANEOUS DIFFERENTIATION 

Compared to LN521, many other feeder-free matrices, such as extracts of the Engelbreth-Holm-Swarm (EHS) tumor, cannot provide the correct biorelevant signals to support hPSCs which often results in loss of pluripotency and genetically drift in vitro. Human PSCs can be cultured to near confluence on LN521, without risk of spontaneous differentiation. The cells remain pluripotent (Oct4+; pink) on LN521 and there is no need to remove differentiated cell areas (DAPI staining only; blue) as compared to cells cultured on other substrates. 

COST-EFFECTIVE HUMAN PSC EXPANSION 

Due to faster growth rate and higher cell yield, the total cost per cell for an average passage is lowest for LN521, compared to other feeder-free matrices. To minimize cost, simply adjust the culture ware size to fit your needs.

Read more about how LN521 saves both time and money for cost-effective pluripotent stem cell culture.

WEEKEND-FREE FEEDING

For reduced labor and cost, the LN521 matrix supports the culture of human PSCs without weekend-free feeding. The protocol is flexible - just feed your cells daily during weekdays and passage your cells on Thursday or Friday. Your hPSC cultures will maintain characteristic cell morphology, high pluripotent cell marker expression, and high expansion rates. Read more about weekend-free hPSC culture protocol.

Download the weekend-free feeding application note. 

EASY AND FLEXIBLE CULTURE PROTOCOLS

When using LN521, your cell culture protocol can easily be made totally defined and animal origin-free, with your choice of culture medium and dissociation reagent.

1. Coat plates with laminin.

2. Wash the cells with PBS and add dissociation reagent of choice - incubate

3. Centrifuge the single-cell suspension and resuspend the pellet in a fresh medium of choice

4. Seed as a single-cell suspension on fresh laminin-coated plates
   
   

Download the coating protocol and the protocol for how to culture hPSCs on LN521.
 

We have visualized the article Monolayer culturing and cloning of human pluripotent stem cells on laminin 521–based matrices under animal origin-free and chemically defined conditions by Rodin et al., published in Nature Protocols 2014. This is a quick guide and overview of how to utilize LN521 as your matrix of choice for your hPSCs through the complete stem cell propagation process.

Here you can order the A1 size hPSC culture poster or download the PDF HERE.

RECOMMENDED PUBLICATIONS

• hPSC Derivation, Maintenance, and Cryopreservation on LN521
• iPSC Reprogramming and Expansion on LN521
• Clonal Culture and Genome Editing on LN521