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LAMININ-521

 

=> GLOMERULAR ENDOTHELIAL CELL
         CULTURE PROTOCOL

A simple, reproducible model of quiescent glomerular endothelial cell monolayer
 

  

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LAMININ KEY ADVANTAGES

  • Laminin-521 enables a simple, reproducible and defined glomerular basement membrane filtration model

  • Laminin-521 promotes all of the characteristics of the quiescent hGEnC monolayer: cobblestone morphology, well-defined adherens junctions and physiological perm-selectivity

  • Culture on laminin resulted in a physiologically relevant barrier

  • High hGEnC attachment and spreading on laminin

  • Laminin-521 also support culture of podocytes

  • Defined and xeno-free culture method for reproduceble experments

 


Renal cell culture applications on human recombinant laminin substratesLaminin-521 culture provides a simple, reproducible and defined glomerular basement membrane filtration model

Laminin-521 is the major laminin expressed in the mature glomerular basement membrane (GBM). However, during glomerulogenesis there are developmental transitions in laminin trimer deposition. Initially the GBM contains laminin-111, laminin-411 and laminin-511. By the capillary loop stage during GMB maturation, laminin-111 is eliminated from the GBM and the beta 2 chain begins to accumulate while the beta 1 chain is down regulated. At maturity, only laminin-521 is detected in the GBM.


Primary glomerular endothelial cells (GEnCs) is an important tool for studying glomerulosclerotic mechanisms and in drug development. In a publication by Pajęcka et al., the authors establish a simple, reproducible and defined in vitro model of quiescent hGEnC monolayer on recombinant human laminin 521 (laminin-521). Laminin-521 best supported hGEnC attachment and spreading and culture on laminin resulted in a physiologically relevant barrier to 70 kDa dextrans which was 82 % tighter than that formed on collagen type IV. Furthermore, only hGEnCs cultured on laminin showed plasma-membrane localized zonula occludens-1 and vascular endothelial cadherin indicative of proper tight and adherens junctions. Laminin-521 promotes all of the characteristics of the quiescent hGEnC monolayer: cobblestone morphology, well-defined adherens junctions and physiological perm-selectivity.


Laminin-521 is the major laminin expressed in the mature glomerular basement membrane and support culture of podocytes

The a-5 chain laminins (laminin-521 and laminin-511) binds predominantly to the integrin α3β1 receptor on the basal surface of podocytes. The binding of laminin-521 and/or laminin-511 to this integrin is essential for the formation of the typical glomerular capillary loop structure. Lack of binding to the α3β1 receptor results in severe kidney glomerular defects during development (Kikkawa et al., 2003). 


Mesangial cells produce laminin-421

Mesangial cells produce laminin-421 and organize glomerular capillaries by binding the laminin α-5 via α3β1 integrin and Lutheran receptor (Kikkawa et al., 2003). Thus, mesangial cells are important for the development and reparation of microvasculature and glomerulogenesis in general (Abrass, 2010). For culture of mesangial cells, we recmmend laminin-421.

Recommended publications

Glomerular endothelial cells and podocytes


Mesangial cells