Integration of embryonic nephrogenic cells carrying a reporter gene into functioning nephrons.
We developed a procedure to introduce and stably express foreign genes into the kidney. The Lac Z reporter gene encoding the bacterial protein beta-galactosidase was introduced by retrovirus-mediated gene transfer into rat nephrogenic mesenchymal cells, which were induced for 24 h with embryonic spinal cord in vitro. The Lac Z-tagged mesenchymal cells were subsequently transplanted underneath the capsule of the neonatal kidney. Two weeks after transplantation, the Lac Z-tagged cells derived from transplants were identified by their beta-galactosidase expression. Well-differentiated Lac Z positive cells were observed in glomerulus and proximal and distal nephron segments. To determine if the tagged mesenchymal cells developed into functional nephrons, fluorescein isothiocyanate-labeled dextran was infused into transplanted animals before death. We observed that fluorescent apical vesicles were colocalized to beta-galactosidase positive proximal tubular cells, indicating that the transplanted mesenchymal cells were integrated into reabsorbing nephrons. These results show the feasibility of introducing foreign genes into epithelia of functioning nephron segments.
Koseki
C
C
Department of Medicine, College of Physicians and Surgeons, Columbia University, New York, New York 10032.
Herzlinger
D
D
al-Awqati
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Q
D000818:Animals; D000831:Animals, Newborn; D002678:Chimera; D003911:Dextrans; D005260:Female; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives*; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006651:Histocytochemistry; D007668:Kidney / Q000196:embryology* / Q000201:enzymology / Q000254:growth & development; D011247:Pregnancy; D051381:Rats; D011919:Rats, Inbred Strains; D014162:Transfection; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics*
D003911:Dextrans; D005452:Fluoresceins; D005456:Fluorescent Dyes; C015219:fluorescein isothiocyanate dextran; D001616:beta-Galactosidase; D016650:Fluorescein-5-isothiocyanate