Document Detail

Immunodissection and culture of rabbit cortical collecting tubule cells.
MedLine Citation:
PMID:  3017126     Owner:  NLM     Status:  MEDLINE    
A mouse monoclonal antibody designated IgG3(rct-30) has been prepared that reacts specifically with an antigen on the surface of all cells comprising the cortical and medullary rabbit renal collecting tubule including the arcades. Plastic culture dishes coated with IgG3(rct-30) were used to isolate collecting tubule cells from collagenase dispersions of rabbit renal cortical cells by immunoadsorption. Typically, 10(6) rabbit cortical collecting tubule (RCCT) cells were obtained from 5 g of renal cortex (2 kidneys). Initial purity was greater than 96% based on immunocytofluorescent staining with three different anti-collecting tubule antibodies. Between 20 and 30% of the RCCT cells were reactive with peanut lectin suggesting that RCCT cells are a mixture of principal and intercalated cells. Approximately 10(7) RCCT cells were obtained after 4 to 5 days in primary culture. Moreover, RCCT cells continued to proliferate after passaging with a doubling time of approximately 32 h. RCCT cells passaged once and then cultured 4-5 days were found 1) to synthesize cAMP in response to arginine vasopressin (AVP), prostaglandin E2 (PGE2), isoproterenol, and parathyroid hormone, but not calcitonin, prostaglandin D2, or prostaglandin I, and 2) to release PGE2 in response to bradykinin but not arginine vasopressin or isoproterenol. Our results indicate that cultured RCCT cells retain many of the hormonal, histochemical, and morphological properties expected for a mixture of principal and intercalated rabbit cortical collecting tubule epithelia. RCCT cells should prove useful both for studying hormonal interactions in the cortical collecting tubule and as a starting population for isolating intercalated collecting tubule epithelia.
W S Spielman; W K Sonnenburg; M L Allen; L J Arend; K Gerozissis; W L Smith
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The American journal of physiology     Volume:  251     ISSN:  0002-9513     ISO Abbreviation:  Am. J. Physiol.     Publication Date:  1986 Aug 
Date Detail:
Created Date:  1986-09-17     Completed Date:  1986-09-17     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0370511     Medline TA:  Am J Physiol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  F348-57     Citation Subset:  IM    
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MeSH Terms
Antibodies, Monoclonal / biosynthesis
Antigens / analysis
Cell Separation
Cells, Cultured
Cyclic AMP / biosynthesis
Cytological Techniques*
Dissection / methods*
Hormones / pharmacology
Immunologic Techniques
Kidney Cortex
Kidney Tubules / cytology*
Kidney Tubules, Collecting / cytology*,  drug effects,  immunology,  metabolism
Prostaglandins E / biosynthesis
Grant Support
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Antigens; 0/Hormones; 0/Prostaglandins E; 363-24-6/Dinoprostone; 60-92-4/Cyclic AMP

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