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LAMININ-521 and LAMININ-511


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PROTOCOL

Maintaining pluripotent mouse ES cells in vitro without addition of leukemia inhibitory factor


CUSTOMER TESTIMONIALS        

HOW TO CULTURE ANIMAL STEM CELLS ON LAMININ SUBSTRATES

How to culture animal stem cells on laminin substratesThe laminin protein family is highly conserved

Laminins are a large family of conserved, multi-domain trimeric basement membrane proteins that contribute to the structure of extracellular matrix and influence the behaviour of associated cells. In the developing embryo, the alpha-1 and alpha-5 laminins are the first extracellular matrix proteins to be expressed. Laminin-111 is mostly expressed in extra-embryonic basement membranes and stimulates cells to commit to differentiation. Contrary, laminin-521 and laminin-511 are expressed around the stem cells in the inner cell mass and support survival and self-renewal of the pluripotent stem cells (Domogatskaya, 2012; Rodin, 2014a). The relative similarity of each pair of laminin chains within a chain constellation is conserved in most vertebrate species (Domogatskaya et al., 2012) and human recombinant laminins has been used for defined and reliable culture of many animal origin pluripotent stem cells, such as mouse, rabbit and monkey.


Culture of Mouse Pluripotent Stem Cells

In contrast to the inner cell mass of the human embryo where both laminin-521 and laminin-511 is expressed, the only laminin isoform expressed in the early mouse embryo is laminin-511. This data is nicely presented in an article published in Nature Cell Biology by the groups of Drs. Austin Smith and Jennifer Nichols where they also show that laminin-511 is crucial for acquisition of naive pluripotency (Boroviak et al., 2014).

Since human recombinant laminin-511 provides the mouse pluripotent stem cells with their natural environment it supports long-term expansion of mouse embryonic stem cells (Domogatskaya et al., 2008). LN-511 alone, without the addition of leukemia inhibitory factor (LIF) or other differentiation inhibitors, is sufficient to enable self-renewal of mouse embryonic stem cells (mESCs) for up to 169 days (31 passages) with maintained expression of pluripotency markers. Cells cultured for 95 days (17 passages) on laminin-511 were used to successfully generate chimeric mice (Domogatskaya et al., 2008). This data is supported by a publication by Lecht and co-workers where they show that laminin is one of the major pro-adhesive ECM protein for mESCs. The mESCs were found to adhere with highest potencies to laminin-511 and laminin-521, most likely via specific binding to a3B1 and a6B1 integrin receptors expressed on the mES cell surface (Lecht et al., 2013).


The main benefits of LN-511 for mouse pluripotent stem cells are:

  • Fast adherence of mESCs to laminin-511
    3 to 5-fold higher affinity than other matrices 
  • Rapid cell expansion with minimal spontaneous differentiation 
    Reliable and fast scale-up where cells grow as a homogenous monolayer 
  • Biorelevant matrix where no differentiation inhibitors are needed
    Self-renewal of mouse ES cells for over 5 months without the presence of LIF 
  • Highly efficient single-cell mESC derivation
    Laminin-511 is crucial for acquisition of naive pluripotency 
  • Chemically defined
    No lot-to-lot variation for less experimental variation
  • Easy coating and single-cell split/seeding protocol
    Standardized monolayer culture system for automation
  • Scientifically proven in high-impact journals
    Validated methods


Culture of Monkey Pluripotent Stem Cells

Laminin-521 and laminin-511 are two of the first extra cellular proteins expressed during development. Both laminin isoforms are present in the inner cell mass of the human embryo and expressed by the pluripotent cells (Domogatskaya et al., 2012). 

Human recombinant laminin-521 is a chemically defined and xeno-free system for robust, long-term expansion of human pluripotent stem cells. Since it successfully replicates the specific human stem cell niche in vitro, it makes it the optimal matrix for efficient generation and robust large-scale expansion of human ES and iPS cells (Rodin et al. 2014). Several researchers are successfully routinely using laminin-521 for pluripotent expansion of monkey pluripotent stem cells (unpublished data).

LAMININ KEY ADVANTAGES

  • Laminins are a major component of the basement membrane that forms the stem cell niche

  • Evident role of laminin-binding integrins in tumorigenesis, both tumour-promoting and suppressive activities

  • Laminin-332 has been shown to be a molecular marker for cells with invasive properties and correlated with poor prognosis

  • Prominent role for laminin-332 as part of the stem cell nice in human hepatic tumours, melanoma cancer and prostate cancer

  • Breast cancer stem cells produce laminin-521 and laminin-511 that promotes self-renewal and tumour initiation via α6β1 integrin interaction and activation of the Hippo transducer TAZ

  • Laminin-521 provide a good support for culture of various cancer cells, such as neuroblastoma, glioblastoma and medulloblastoma cells

  • Laminin-521 have specific effects in promoting the stemness of glioma cells, both at gene and protein expression level

  • Defined and xeno-free cell culture matrix for clinical compliance

  • No lot-to-lot variability for standardized experiments with less variation

  • Scientifically validated in high-impact journals

  • x
  • Expansion of human PSC

  • Clonal cell culture applications

  • Eye cells

  • Cardiac cells

  • Neural cells

  • Skeletal muscle cells

  • Kidney cells

  • Hepatic cells

  • Cancer cells

  • Lung cells

  • Intestinal cells

  • Pancreatic cells

  • Hematopoietic stem and progenitor cells

  • Mesenchymal stem cells

  • Animal stem cells

  • Endothelial cells

  • Normal and cancerous mammary cells

  • Epithelial cells

Laminin-521

Laminin-521 human stem cell matrix makes pluripotent stem cell culture easy in a defined, xeno-free and biologically relevant cell culture system.

  • x
  • Neural cells

  • Hepatic cells

  • Lung cells

  • Intestinal cells

  • Normal and cancerous mammary cells

  • Expansion of human PSC

  • Clonal cell culture applications

  • Eye cells

  • Cardiac cells

  • Skeletal muscle cells

  • Kidney cells

  • Cancer cells

  • Pancreatic cells

  • Hematopoietic stem and progenitor cells

  • Mesenchymal stem cells

  • Animal stem cells

  • Endothelial cells

  • Epithelial cells

Laminin-111

Laminin-111 is important during early epithelial development and differentiation of the epiblast, and is commonly used as a general attachment protein in cell culture.

  • x
  • Neural cells

  • Eye cells

  • Endothelial cells

  • Animal stem cells

  • Pancreatic cells

  • Epithelial cells

  • Cardiac cells

  • Cancer cells

  • Hepatic cells

  • Intestinal cells

  • Kidney cells

  • Lung cells

  • Skeletal muscle cells

  • Expansion of human PSC

  • Clonal cell culture applications

  • Hematopoietic stem and progenitor cells

  • Mesenchymal stem cells

  • Normal and cancerous mammary cells

LAMscreen

LAMscreen is a kit with all our currently available laminin isoforms so that you can easily screen the best laminins for different cell types.