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.jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117 J+Am+Soc+Nephrol
2014 ; 25
(6
): 1211-25
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Rapid and efficient differentiation of human pluripotent stem cells into
intermediate mesoderm that forms tubules expressing kidney proximal tubular
markers
#MMPMID24357672
Lam AQ
; Freedman BS
; Morizane R
; Lerou PH
; Valerius MT
; Bonventre JV
J Am Soc Nephrol
2014[Jun]; 25
(6
): 1211-25
PMID24357672
show ga
Human pluripotent stem cells (hPSCs) can generate a diversity of cell types, but
few methods have been developed to derive cells of the kidney lineage. Here, we
report a highly efficient system for differentiating human embryonic stem cells
and induced pluripotent stem cells (referred to collectively as hPSCs) into cells
expressing markers of the intermediate mesoderm (IM) that subsequently form
tubule-like structures. Treatment of hPSCs with the glycogen synthase kinase-3?
inhibitor CHIR99021 induced BRACHYURY(+)MIXL1(+) mesendoderm differentiation with
nearly 100% efficiency. In the absence of additional exogenous factors,
CHIR99021-induced mesendodermal cells preferentially differentiated into cells
expressing markers of lateral plate mesoderm with minimal IM differentiation.
However, the sequential treatment of hPSCs with CHIR99021 followed by fibroblast
growth factor-2 and retinoic acid generated PAX2(+)LHX1(+) cells with 70%-80%
efficiency after 3 days of differentiation. Upon growth factor withdrawal, these
PAX2(+)LHX1(+) cells gave rise to apically ciliated tubular structures that
coexpressed the proximal tubule markers Lotus tetragonolobus lectin, N-cadherin,
and kidney-specific protein and partially integrated into embryonic kidney
explant cultures. With the addition of FGF9 and activin, PAX2(+)LHX1(+) cells
specifically differentiated into cells expressing SIX2, SALL1, and WT1, markers
of cap mesenchyme nephron progenitor cells. Our findings demonstrate the
effective role of fibroblast growth factor signaling in inducing IM
differentiation in hPSCs and establish the most rapid and efficient system
whereby hPSCs can be differentiated into cells with features characteristic of
kidney lineage cells.