title: Luminescence-based assay to screen preservation solutions for optimal ability to maintain viability of rat intestinal grafts.
abstract: BACKGROUND
  Segmental intestinal transplantations from living, genetically related donors provide advantages compared with those from cadaveric subjects. However, successful preservation during ischemic cold storage is critical for living donor grafts. Thus, the development of preservation solutions that maintain graft viability is essential for success. Herein we have reported application of a cell-based viability assay in multiwell plates to assess the effectiveness of various solutions to preserve intestinal grafts.
  
  
  METHODS
  Freshly isolated intestinal chips from luciferase transgenic rats were placed in 96-well tissue culture plates for incubation at 4°C for 24 hours in various preservation solutions: ET-Kyoto (ET-K), University of Wisconsin (UW) solution, Euro-Collins (EC) solution, histidine-tryptophan-ketoglutarate (HTK) solution, lactated Ringer's (LR) solution, or saline.
  
  
  RESULTS
  As indicated by a higher level of luminescence, intestinal chips preserved in UW, HTK, or ET-K solution contained more viable cells, than those preserved in EC, LR, or saline solution. After exposure to the preservation solutions for 1 hour, the mucosal layer chips showed lower cell viability than the muscle layer chips.
  
  
  CONCLUSIONS
  Our data demonstrated that ET-K and UW solutions used together with intestinal chips of Luciferase transgenic rat and in vivo imaging provided optimal viability during ischemic cold storage prior to transplantation. Further development of preservation conditions to minimize the loss of viability of intestinal grafts before clinical transplantation is essential to improve outcomes.
authors: Division of Development of Advanced Treatment Center for Development of Advanced Medical Technology, Jichi Medical University, Shimotsukeshi, Tochigi, Japan.
authors: N
authors: N
authors: Kasahara
authors: T
authors: T
authors: Kikuchi
authors: J
authors: J
authors: Doi
authors: T
authors: T
authors: Teratani
authors: Y
authors: Y
authors: Fujimoto
authors: S
authors: S
authors: Uemoto
authors: Y
authors: Y
authors: Yasuda
authors: E
authors: E
authors: Kobayashi
mesh_terms: D000241:Adenosine / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism; D000493:Allopurinol / Q000494:pharmacology; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D050377:Cold Ischemia / Q000009:adverse effects; D005942:Gluconates / Q000494:pharmacology; D005947:Glucose / Q000494:pharmacology; D005978:Glutathione / Q000494:pharmacology; D057166:High-Throughput Screening Assays / Q000379:methods*; D006895:Hydroxyethyl Starch Derivatives / Q000494:pharmacology; D006982:Hypertonic Solutions / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D007421:Intestine, Small / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology / Q000637:transplantation*; D007552:Isotonic Solutions / Q000494:pharmacology; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D008163:Luminescent Measurements; D008353:Mannitol / Q000494:pharmacology; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions / Q000494:pharmacology*; D010710:Phosphates / Q000494:pharmacology; D011189:Potassium Chloride / Q000494:pharmacology; D011343:Procaine / Q000494:pharmacology; D011887:Raffinose / Q000494:pharmacology; D051381:Rats; D055647:Rats, Transgenic; D000077325:Ringer's Lactate; D012965:Sodium Chloride / Q000494:pharmacology; D013050:Spectrometry, Fluorescence; D013997:Time Factors; D046509:Tissue Culture Techniques; D014199:Trehalose / Q000494:pharmacology
chemical_list: C039984:Bretschneider cardioplegic solution; C087059:ET-Kyoto solution; C037096:Euro-Collins' solution; D005942:Gluconates; D006895:Hydroxyethyl Starch Derivatives; D006982:Hypertonic Solutions; D007328:Insulin; D007552:Isotonic Solutions; D019188:Organ Preservation Solutions; D010710:Phosphates; D000077325:Ringer's Lactate; C055206:University of Wisconsin-lactobionate solution; D008353:Mannitol; D012965:Sodium Chloride; D011343:Procaine; D000493:Allopurinol; D011189:Potassium Chloride; D000255:Adenosine Triphosphate; D014199:Trehalose; D008156:Luciferases; D005978:Glutathione; D005947:Glucose; D000241:Adenosine; D011887:Raffinose