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PRODUCT RECOMMENDATION

LAMININ-521 

 
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PROTOCOL

Effective production of hESC-RPE cells in a xeno-free and defined manner


Production and transplantation of GMP grade xeno-free hESC-derived RPE cells


VEIW ON DEMAND

Defined and xeno-free differentiation of clinically compliant hESC-RPE cells

GMP-compliant stem cell differentiation to dopaminergic neurons on laminin


APPLICATION NOTE

Get the hPSC culture application note here


ARTICLES

LAMININ KEY ADVANTAGES

  • Defined and xeno-free differentiation method for clinically compliant hESC-RPE

  • Laminin-521 supports high seeding efficiency and cell migration

  • Laminin-521 cultured hESC-RPE cells exhibit native characteristics including morphology, marker expression, monolayer integrity, pigmentation, polarization and phagocytic activity

  • Cells transplanted in suspension in a large-eyed animal model incorporate as a nicely polarized monolayer

  • Transplanted cells show long-term, in vivo functionality, including phagocytic activity and photoreceptor rescue

  • Laminin culture substrates is a part of FDA approved IND, and a Phase 1/2a clinical trial for AMD patients is currently ongoing.

  • Main isoforms expressed in Bruch’s membrane are laminin-521, laminin-511, laminin-332 and laminin-111

  • A biologically relevant culture environment for human ES and iPS cells

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

 


 Effective generation of hESC-RPE cells under defined and xeno-free conditions

In a publication by the groups of Drs. Hovatta, Lanner and Kvanta, the authors describe a protocol for effective differentiation of hESC-RPE cells on laminin-521 in a xeno-free and defined manner (Plaza Reyes, 2015). The differentiated RPE cells exhibit native characteristics including morphology, pigmentation, marker expression, monolayer integrity, polarization and phagocytic activity. The authors also established a large-eyed geographic atrophy model that allow imaging of the hESC-RPE and host retina. Cells are transplanted in suspension and show long-term integration and form polarized monolayers exhibiting phagocytic and photoreceptor rescue capacity.


In a recent publication, researchers at Cell Cure Neurosciences Ltd. display the efficacy of RPE cells derived under xeno-free conditions from clinical and xeno-free grade human embryonic stem cells following transplantation into the subretinal space of Royal College of Surgeons (RCS) rats (McGill, 2017). Again, laminin cell culture substrate is being used in the differentiation protocol. The results demonstrate that transplantation of hESC-RPE cells into the subretinal space of RCS rats protected retinal structure, rescued visual function, preserved rod and cone photoreceptors long-term. OKT was rescued in a dose-dependent manner and outer nuclear layer (ONL) was significantly thicker in cell-treated eyes than controls. This data combined with data collected in a definitive safety studies (tumorigenicity and spiking and safety/biodistribution) has resulted in an FDA approved IND (investigational new drug) and a Phase 1/2a clinical trial for AMD patients is currently ongoing, NCT02286089.


Bruch’s membrane, contains several laminins isoforms

It has been shown that the beta-2 chain of the laminin molecule is important for the RPE cells (Libby, 1996). It surrounds RPE cells at the onset of rod photoreceptor birth and is present on the apical surface of the retinal neuroepithelium. At birth, laminin that contain the beta-2 chain fills the matrix between the juxtaposed surfaces of the RPE and neuroepithelium. It is present throughout postnatal development and is also is found in association with blood vessels in the neural retina. A publication by Aisenbrey and colleagues show that the multilayered extracellular matrix underlying the retina, Bruch’s membrane, contains several laminins that retinal pigment epithelial cells grow on. The main isoforms expressed are laminin-521, laminin-511, laminin-332 and laminin-111. RPE cells adhere robustly to all laminins of Bruch´s membrane and actively synthesize these laminins (Aisenbrey, 2006).

RECOMMENDED PUBLICATIONS

  • 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.