Highly Specific Separation of Heterogeneous Cell Populations by Lectin-Coated Beads: Application for the Isolation of Inner Medullary Collecting Duct CellsGrupp C.a · Troche I.a · Steffgen J.a · Langhans S.b · Cohen D.I.b · Brandl L.b · Müller G.A.a
a Abteilung Nephrologie und Rheumatologie, Zentrum Innere Medizin, Georg-August-Universität, Göttingen, und b Sektion Nephrologie, Medizinische Klinik, Universität Ulm, Deutschland
Do you have an account?
- Rent for 48h to view
- Buy Cloud Access for unlimited viewing via different devices
- Synchronizing in the ReadCube Cloud
- Printing and saving restrictions apply
Rental: USD 8.50
Cloud: USD 20.00
Conditions for the highly specific selection of a cell type by the use of lectin-coated magnetic beads are reported for the isolation of inner medullary collecting duct (IMCD) cells from a heterogeneous inner medullary cell suspension, containing both single cells and tubular fragments of variable size. The lectin Dolichos Biflorus Agglutinin (DBA), which binds in rat inner medulla exclusively to IMCD cells, was coupled via the avidin-biotin system to beads. By isolating DBA-bead-IMCD cells in a magnetic field (positive selection) from a suspension containing about 50% IMCD, a fraction of 98 ± 1% purity was obtained; recovery of cells was up to 90%. Suspensions negative on reverse-transcriptase polymerase chain reaction for vimentin as a marker of contaminating interstitial and vascular cells could be received by repeating this procedure and additional trypsinization. On the other hand, it was possible to reduce the portion of IMCD cells in the suspension by one isolation step to 1.5 ± 0.9% (negative selection). Performing this step twice resulted in virtually pure suspensions. No significant effects of this isolation technique on cell viability, growth characteristics, and biochemical parameters were observed. Therefore, this method appears to be a powerful tool for the highly specific separation of heterogeneous cell populations.
- Stokes JB, Grupp C, Kinne RKH: Purification of rat papillary collecting duct cells: Functional and metabolic assessment. Am J Physiol 1987;253:F251–F262.
- Grenier FC, Rollins TE, Smith WL: Kinin-induced prostaglandin synthesis by renal papillary collecting tubule cells in culture. Am J Physiol 1981;241:F94–104.
- Heidrich HG, Hannig K: Separation of cell populations by free-flow electrophoresis. Methods Enzymol 1989;171:513–531.
- Orfao A, Ruiz-Arguelles A: General concepts about cell sorting techniques. Clin Biochem 1996;29:5–9.
- Gaudernack G, Leivestad T, Ugelstad J, Thorsb E: Isolation of pure functionally active CD8+ T cells: Positive selection with monoclonal antibodies directly conjugated to monosized magnetic microspheres. J Immunol Methods 1986;90:179–187.
- Lundkvist A, Horling J, Athlin L, Rosen A, Niklasson B: Neutralizing human monoclonal antibodies against Puumala virus, causative agent of nephropathia epidemica: A novel method using antigen-coated magnetic beads for specific B cell isolation. J Gen Virol 1993;74:1303–1310.
- Schmitz B, Radbruch A, Kummel T, Wickenhauser C, Korb H, Hansmann ML, Thiele J, Fischer R: Magnetic activated cell sorting (MACS) – a new immunomagnetic method for megakaryocytic cell isolation: Comparison of different separation techniques. Eur J Haematol 1994;52:267–275.
- Raghunath M, Grupp C, Neumann I, Heidtmann A, Roelcke D: Polylactosamine sugar chains expressed by epithelia of Henle’s loop and collecting duct in rat and human kidney are selectively recognized by human cold agglutinins anti-I/i. Tissue Antigens 1994;44:159–165.
- Grupp C, Lottermoser J, Cohen DI, Begher M, Franz HE, Muller GA: Transformation of rat inner medullary fibroblasts to myofibroblasts in vitro. Kidney Int 1997;52:1279–1290.
- Grupp C, Begher M, Cohen D, Raghuanth M, Franz HE, Muller GA: Isolation and characterization of the lower portion of the thin limb of Henle in primary culture. Am J Physiol 1998;274:F775–F782.
- Terada Y, Tomita K, Nonoguchi H, Yang T, Marumo F: Different localization and regulation of two types of vasopressin receptor messenger RNA in microdissected rat nephron segments using reverse transcription polymerase chain reaction. J Clin Invest 1993;92:2339–2345.
- Yamamoto T, Feng L, Mizuno T, Hirose S, Kawasaki K, Yaoita E, Kihara I, Wilson CB: Expression of mRNA for natriuretic peptide receptor subtypes in bovine kidney. Am J Physiol 1994;267:F318–F324.
- Zhuo J, Alcorn D, Allen AM, Mendelsohn FA: High resolution localization of angiotensin II receptors in rat renal medulla. Kidney Int 1992;42:1372–1380.
- Chow LH, Subramanian S, Nuovo GJ, Miller F, Nord EP: Endothelin receptor mRNA expression in renal medulla identified by in situ RT-PCR. Am J Physiol 1995;269:F449–F457.
- Huo T, Ye MQ, Healy DP: Characterization of a dopamine receptor (DA2K) in the kidney inner medulla. Proc Natl Acad Sci USA 1991;88:3170–3174.
- Teitelbaum I: Hormone signaling systems in inner medullary collecting ducts. Am J Physiol 1992;263:F985–F990.
- Bonvalet JP, Pradelles P, Farman N: Segmental synthesis and actions of prostaglandins along the nephron. Am J Physiol 1987;253:F377–F387.
- Kohan DE, Hughes AK: Autocrine role of endothelin in rat IMCD: Inhibition of AVP-induced cAMP accumulation. Am J Physiol 1993;265:F126–F129.
- Salido EC, Lakshmanan J, Fisher DA, Shapiro LJ, Barajas L: Expression of epidermal growth factor in the rat kidney: An immunocytochemical and in situ hybridization study. Histochemistry 1991;96:65–72.
- Pavenstadt-Grupp I, Grupp C, Kinne RK: Measurement of element content in isolated papillary collecting duct cells by electron probe microanalysis. Pflügers Arch 1989;413:378–384.
- Labarca C, Paigen K: A simple, rapid, and sensitive DNA assay procedure. Anal Biochem 1980;102:344–352.
- Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193:265–275.
- Lemley KV, Kriz W: Anatomy of the renal interstitium. Kidney Int 1991;39:370–381.
- Clapp WL, Madsen KK, Verlander JL, Tisher CC: Intercalated cells of the rat inner medullary collecting duct. Kidney Int 1987;31:1080–1087.
Treleaven JG, Gibson FM, Ugelstad J, Rembaum A, Philip T, Caine GD, Kemshead JT: Removal of neuroblastoma cells from bone marrow with monoclonal antibodies conjugated to magnetic microspheres. Lancet 1984;i:70–73.
- Pilling D, Kitas GD, Salmon M, Bacon PA: The kinetics of interaction between lymphocytes and magnetic polymer particles. J Immunol Methods 1989;122:235–241.
- Van Adelsberg J, Edwards JC, Herzlinger D, Cannon C, Rater M, Al Awqati Q: Isolation and culture of HCO3-secreting intercalated cells. Am J Physiol 1989;256:C1004–C1011.
- Holthofer H: Lectin binding sites in kidney: A comparative study of 14 animal species. J Histochem Cytochem 1983;31:531–537.
- Le Hir M, Dubach UC: The cellular specificity of lectin binding in the kidney. II. A light microscopical study in the rabbit. Histochemistry 1982;74:531–540.
- Silva FG, Nadasdy T, Laszik Z: Immunohistochemical and lectin dissection of the human nephron in health and disease. Arch Pathol Lab Med 1993;117:1233–1239.
- Aguirre JI, Han JS, Itagaki S, Doi K: Lectin histochemical study on the kidney of normal and streptozotocin-induced diabetic hamsters. Histol Histopathol 1993;8:273–278.
Article / Publication Details
Copyright / Drug Dosage / DisclaimerCopyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.