Publications

A Biomimetic Electrospun Membrane Supports the Differentiation and Maturation of Kidney Epithelium from Human Stem Cells

Mou, X., Shah, J., Bhattacharya, R., Kalejaiye, T.D., Sun, B., Hsu, P.-C., Musah, S Bioengineering, 2022

Laminin (Biolaminin 511) was shown to be crucial for the differentiation of hPSCs into mature kidney glomerular podocytes on electrospun silk fibroin. Without laminin functionalization, the cells were poorly attached and scarce having small and rounded morphology. Electrospun silk fibroin membranes are biomimetic scaffolds, mimicking the surface topography of the glomerulus. These kinds of biomaterials can be integrated into microphysiological systems, such as microfluidic organ-on-chip devices or co-culture systems for modelling tissue development and disease.

ECM modulated early kidney development in embryonic organ culture

Sebinger D., Ofenbauer A., Gruber P., Malik S., Werner C.Biomaterials, 2013

Here, the authors, explore the role of the extracellular matrix (ECM) constituents on renal structure formation during renal organogenesis. By culture of microdissected E11.5 kidney rudiments, the show that specific ECM components, including collagen I and laminin, supported nephronal and tubular structure formation of the developing organ. Culture on laminin-111 promoted development as shown by more bud tips and more nephrons.

Laminin isoforms and their receptors in the developing kidney

Ekblom P., Klein G., Ekblom M., Sorokin L.Am J Kidney Dis., 1991

Epithelial cells have a polarized morphology, with distinct basal, lateral, and apical cell surfaces. It would be of considerable interest to know how the polarized morphology develops during embryogenesis. Both the tubular and glomerular epithelial cells of the kidney develop from mesenchymal stem cells during embryogenesis. Unique conversion of nonpolar cells to polarized epithelial cells thus occurs in the embryonic kidney. This conversion also occurs in vitro if the mesenchymal cells are properly induced. Organ cultures of mesenchymal cells from the mouse embryonic kidney have therefore been much used to study the development of epithelial cell polarity. We have used this model system to study the role of basement membrane glycoproteins in development. The results obtained suggest that laminins are particularly important for epithelial cell development. There are many different types of laminins. Developing kidney tubule cells synthesize that promote development by interacting with specific integrin receptors on the cell surface. The mesenchymal stem cells also produce laminin.

Laminin a4-Null Mutant Mice Develop Chronic Kidney Disease with Persistent Overexpression of Platelet-Derived Growth Factor

Abrass C.K., Hansen K.M., Patton B.L.Matrix Pathobiology, 2010

Laminin-8/9 is synthesized by rat glomerular mesangial cells and is required for PDGF-induced mesangial cell migration. Here the authors show that Lama4-/- mice have progressive glomerular and tubulointerstitial fibrosis. These mice have a significant increase in the expression of platelet-derived growth factor (PDGF)-BB, PDGF-DD, and PDGF receptor beta in association with immature glomerular and peritubular capillaries. In addition, mesangial cell exposure to alpha4-containing laminins results in the down-regulation of PDGF receptor mRNA and protein, suggesting a direct effect of LN411/LN421 on vessel maturation. These data suggest that the failure of laminin alpha4-mediated down-regulation of PDGF activity contributes to the progressive renal lesions in this animal model. 

Directed differentiation of human induced pluripotent stem cells into mature kidney podocytes and establishment of a Glomerulus Chip

Musah S., Dimitrakakis N., Camacho D.M., Church G.M., Ingber D.E. Nature Protocols, 2018

With the use of laminin 511 coating, the authors have developed a detailed protocol for the directed differentiation of human iPS cells into mature, post-mitotic kidney glomerular podocytes with high (>90%) efficiency within 26 d and under chemically defined conditions, without genetic manipulations or subpopulation selection. They also describe how these iPS cell-derived podocytes may be induced to form within a microfluidic organ-on-a-chip culture device to build a human kidney Glomerulus Chip that mimics the structure and function of the kidney glomerular capillary wall in vitro.

Glomerular endothelial cells and podocytes jointly synthesize laminin-1 and -11 chains

St John P.L. and Abrahamson D.R.Kidney International, 2001

During the glomerular basement membrane assembly, laminin-111 is initially expressed in vascular clefts of comma- ann S-shaped bodies and is eventually replaced by laminin-521 which persists into maturation. Post-fixation immunoelectron microscopy of developing mouse and 2-3 days old mice kidney was performed and showed Intracellular labeling for laminin-111 and laminin-521 in developing glomerular endothelial cells and podocytes. In early capillary loop stage, GBMs laminin-111 was absent but the expression of a5 was strong (developmental switch). The results show that both endothelial cells and podocytes synthesize laminin but the highest level of laminin synthesis by endothelial cells.

Roles for Laminin in Embryogenesis: Exencephaly, Syndactyly, and Placentopathy in Mice Lacking the Laminin α5 Chain

Miner J.H., Cunningham J., Sanes J.R. J Cell Biol., 1998

Here, we show that the laminin α5 chain is required during embryogenesis. The α5 chain is present in virtually all BLs of early somite stage embryos and then becomes restricted to specific BLs as development proceeds. BLs that lose α5 retain or acquire other α chains. Embryos lacking laminin α5 die late in embryogenesis. They exhibit multiple developmental defects, including failure of anterior neural tube closure. In addition to exencephaly, syndactyly, and placentopathy, several defects were seen in internal organs of Lama5 −/− embryos, including small or absent kidneys. Some of these defects were variable in severity and penetrance, but all can be associated with BLs that normally contain laminin α5.

The Laminin a Chains: Expression, Developmental Transitions, and Chromosomal Locations of a 1-5, Identification of Heterotrimeric Laminins 8–11, and Cloning of a Novel a 3 Isoform

Miner J.H., Patton B.L., Lentz S.I., Gilbert D.J., Snider W.D., Jenkins N.A., Copeland N.G., Sane J.R. The Journal of Cell Biology, 1997

Using a panel of nucleotide and antibody probes, we surveyed the expression of a 1-5 in murine tissues. All five chains were expressed in both embryos and adults, but each was distributed in a distinct pattern at both RNA and protein levels. Overall, a 4 and a 5 exhibited the broadest patterns of expression, while the expression of a1 was the most restricted. Detailed analysis of developing kidney revealed that some individual basal laminae, including those of the tubule and glomerulus, changed in laminin chain composition as they matured. 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.

The glomerular basement membrane

Miner JH. Exp Cell Res. 2012

This review describes the role of glomerular basement membrane (GBM) in the kidney. The GBM consists mainly of laminin (mostly laminin-521), type IV collagen, nidogen, and heparan sulfate proteoglycan.

Synergistic activities of alpha3 and alpha6 integrins are required during apical ectodermal ridge formation and organogenesis in the mouse

Both integrin α6-null mice α3-null mice die at birth, with kidney and lung defects at late stages of development, and skin blistering. To investigate possible overlapping functions between α3 and α6 integrins, we analyzed the phenotype of compound α3−/− and α6−/− mutant embryos. Double homozygous mutant embryos were growth-retarded and displayed several developmental defects not observed in the single mutant animals. The presence of novel phenotypes in double mutant embryos demonstrates the synergism between α3 and α6 integrins and their essential roles in multiple processes during embryogenesis.