PMID- 33719571
OWN - NLM
STAT- MEDLINE
DCOM- 20210805
LR  - 20210805
IS  - 1522-1466 (Electronic)
IS  - 1522-1466 (Linking)
VI  - 320
IP  - 5
DP  - 2021 May 1
TI  - Machine-perfused donor kidneys as a source of human renal endothelial cells.
PG  - F947-F962
LID - 10.1152/ajprenal.00541.2020 [doi]
AB  - Renal endothelial cells (ECs) play crucial roles in vasorelaxation, 
      ultrafiltration, and selective transport of electrolytes and water, but also in 
      leakage of the glomerular filtration barrier and inflammatory processes like 
      complement activation and leukocyte recruitment. In addition, they are target 
      cells for both cellular and antibody-mediated rejection in the transplanted 
      kidney. To study the molecular and cellular processes underlying EC behavior in 
      renal disease, well-characterized primary renal ECs are indispensible. In this 
      report, we describe a straightforward procedure to isolate ECs from the perfusion 
      fluid of human donor kidneys by a combination of negative selection of 
      monocytes/macrophages, positive selection by CD31 Dynabeads, and propagation in 
      endothelium-specific culture medium. Thus, we isolated and propagated renal ECs 
      from 102 donor kidneys, representative of all blood groups and major human 
      leukocyte antigen (HLA) class I and II antigens. The obtained ECs were positive 
      for CD31 and von Willebrand factor, expressed other endothelial markers such as 
      CD34, VEGF receptor-2, TIE2, and plasmalemmal vesicle associated protein-1 to a 
      variable extent, and were negative for the monocyte marker CD14 and lymphatic 
      endothelial marker podoplanin. HLA class II was either constitutively expressed 
      or could be induced by interferon-gamma. Furthermore, as a proof of principle, we 
      showed the diagnostic value of this renal endothelial biobank in renal 
      endothelium-specific cross-matching tests for HLA antibodies.NEW & NOTEWORTHY We 
      describe a new and widely accessible approach to obtain human primary renal 
      endothelial cells in a standardized fashion, by isolating from the perfusate of 
      machine-perfused donor kidneys. Characterization of the cells showed a mixed 
      population originating from different compartments of the kidney. As a proof of 
      principle, we demonstrated a possible diagnostic application in an 
      endothelium-specific cross-match. Next to transplantation, we foresee further 
      applications in the field renal endothelial research.
FAU - Lammerts, Rosa G M
AU  - Lammerts RGM
AUID- ORCID: 0000-0002-9970-9006
AD  - Division of Nephrology, University Medical Center Groningen, University of 
      Groningen, Groningen, The Netherlands.
FAU - Lagendijk, Lisanne M
AU  - Lagendijk LM
AD  - Division of Nephrology, University Medical Center Groningen, University of 
      Groningen, Groningen, The Netherlands.
FAU - Tiller, Gesa
AU  - Tiller G
AD  - Division of Nephrology, University Medical Center Groningen, University of 
      Groningen, Groningen, The Netherlands.
FAU - Dam, Wendy A
AU  - Dam WA
AD  - Division of Nephrology, University Medical Center Groningen, University of 
      Groningen, Groningen, The Netherlands.
FAU - Lancaster, Harriet L
AU  - Lancaster HL
AD  - Division of Nephrology, University Medical Center Groningen, University of 
      Groningen, Groningen, The Netherlands.
FAU - Daha, Mohamed R
AU  - Daha MR
AD  - Division of Nephrology, University Medical Center Groningen, University of 
      Groningen, Groningen, The Netherlands.
FAU - Seelen, Marc A
AU  - Seelen MA
AD  - Division of Nephrology, University Medical Center Groningen, University of 
      Groningen, Groningen, The Netherlands.
FAU - Hepkema, Bouke G
AU  - Hepkema BG
AD  - Transplantation Immunology, Department of Laboratory Medicine, University of 
      Groningen, University Medical Center Groningen, Groningen, The Netherlands.
FAU - Pol, Robert A
AU  - Pol RA
AD  - Department of Surgery, University Medical Center Groningen, University of 
      Groningen, Groningen, The Netherlands.
FAU - Leuvenink, Henri G D
AU  - Leuvenink HGD
AD  - Department of Surgery, University Medical Center Groningen, University of 
      Groningen, Groningen, The Netherlands.
FAU - Molema, Grietje
AU  - Molema G
AD  - Medical Biology Section, Department of Pathology and Medical Biology, University 
      Medical Center Groningen, University of Groningen, The Netherlands.
FAU - van den Born, Jacob
AU  - van den Born J
AD  - Division of Nephrology, University Medical Center Groningen, University of 
      Groningen, Groningen, The Netherlands.
FAU - Berger, Stefan P
AU  - Berger SP
AD  - Division of Nephrology, University Medical Center Groningen, University of 
      Groningen, Groningen, The Netherlands.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20210315
PL  - United States
TA  - Am J Physiol Renal Physiol
JT  - American journal of physiology. Renal physiology
JID - 100901990
RN  - 0 (Histocompatibility Antigens Class I)
SB  - IM
MH  - Cell Separation/*methods
MH  - Cells, Cultured
MH  - Endothelial Cells/*physiology
MH  - Histocompatibility Antigens Class I
MH  - Humans
MH  - Kidney/*blood supply/*cytology
MH  - Organ Culture Techniques/*methods
MH  - Tissue Donors
OTO - NOTNLM
OT  - donation
OT  - endothelial cells
OT  - kidney
OT  - machine perfusion
OT  - perfusate
OT  - transplantation
EDAT- 2021/03/16 06:00
MHDA- 2021/08/06 06:00
CRDT- 2021/03/15 12:32
PHST- 2021/03/16 06:00 [pubmed]
PHST- 2021/08/06 06:00 [medline]
PHST- 2021/03/15 12:32 [entrez]
AID - 10.1152/ajprenal.00541.2020 [doi]
PST - ppublish
SO  - Am J Physiol Renal Physiol. 2021 May 1;320(5):F947-F962. doi: 
      10.1152/ajprenal.00541.2020. Epub 2021 Mar 15.