PMID- 19794906
OWN - NLM
STAT- MEDLINE
DCOM- 20110128
LR  - 20220410
IS  - 1942-0994 (Electronic)
IS  - 1942-0900 (Print)
IS  - 1942-0994 (Linking)
VI  - 1
IP  - 1
DP  - 2008 Oct-Dec
TI  - Primary mouse renal tubular epithelial cells have variable injury tolerance to 
      ischemic and chemical mediators of oxidative stress.
PG  - 33-8
AB  - We have developed and evaluated an in vitro culture method for assessing ischemic 
      injury in primary mouse renal tubular epithelial cells (RTEC) in which to explore 
      the pathobiology underlying acute kidney injury. RTEC were predominately of 
      proximal tubule origin which is most susceptible to ischemic injury as compared 
      to other nephron segments. Oxidative stress was induced by chemically depleting 
      ATP using Antimycin A and 2-Deoxy-D-Glucose and by exposing cells to a 1% oxygen 
      environment. Necrotic injury was assessed by measuring LDH released into culture 
      supernatants. Optimal dose and time of exposure to each injury agent was 
      determined for induction of mild, moderate and severe ischemic injury defined as 
      LDH release of </= 20%, 21-49% and >/= 50% above baseline respectively. Antimycin 
      A and 2-Deoxy-D-Glucose produced a progressive increase in LDH release which was 
      time dependent but chemical concentration independent. A 1% oxygen environment 
      also induced cell injury over time but only if glucose was absent from the 
      culture media. Antimycin A was most effective at inducing oxidative stress 
      causing a mean LDH release of 61% at 48 hr compared to 19% and 50% LDH release 
      induced by 2-Deoxy-D-Glucose and by exposure to 1% oxygen respectively at the 
      same 48 hour time point.The cell culture method described provides several 
      advantages including the use of serum free media and the ability to grow primary 
      cells without matrix support. The LDH assay for injury assessment is 
      reproducible, cost effective, objective and minimizes background cell death. A 
      simple method for the culture and injury of primary mouse renal tubular 
      epithelial cells has thereby been established and provides a useful tool for 
      future investigations of ischemic kidney injury.
FAU - Breggia, Anne C
AU  - Breggia AC
AD  - Maine Medical Center Research Institute, Clinical and Translational Research, 
      Portland, Maine 04102, USA.
FAU - Himmelfarb, Jonathan
AU  - Himmelfarb J
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Oxid Med Cell Longev
JT  - Oxidative medicine and cellular longevity
JID - 101479826
RN  - 642-15-9 (Antimycin A)
RN  - 8L70Q75FXE (Adenosine Triphosphate)
RN  - 9G2MP84A8W (Deoxyglucose)
RN  - EC 1.1.1.27 (L-Lactate Dehydrogenase)
SB  - IM
MH  - Acute Kidney Injury/metabolism/*pathology
MH  - Adenosine Triphosphate/deficiency/metabolism
MH  - Animals
MH  - Antimycin A/pharmacology
MH  - Cell Death/physiology
MH  - Cell Hypoxia/physiology
MH  - Cells, Cultured
MH  - Deoxyglucose/pharmacology
MH  - Epithelial Cells/metabolism/pathology
MH  - Ischemia/metabolism/pathology
MH  - Kidney Tubules, Proximal/blood supply/*metabolism/*pathology
MH  - L-Lactate Dehydrogenase/metabolism
MH  - Male
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Oxidative Stress/physiology
PMC - PMC2715195
OTO - NOTNLM
OT  - ATP
OT  - acute kidney injury
OT  - glycolysis
OT  - ischemia
OT  - lactate dehydrogenase
OT  - necrosis
OT  - renal tubular epithelial cells
EDAT- 2009/10/02 06:00
MHDA- 2011/02/01 06:00
CRDT- 2009/10/02 06:00
PHST- 2008/05/14 00:00 [received]
PHST- 2008/06/19 00:00 [revised]
PHST- 2008/06/23 00:00 [accepted]
PHST- 2009/10/02 06:00 [entrez]
PHST- 2009/10/02 06:00 [pubmed]
PHST- 2011/02/01 06:00 [medline]
AID - 1942-0900-1-1-4 [pii]
AID - 10.4161/oxim.1.1.6491 [doi]
PST - ppublish
SO  - Oxid Med Cell Longev. 2008 Oct-Dec;1(1):33-8. doi: 10.4161/oxim.1.1.6491.