A luminance-based heart chip assay for assessing the efficacy of graft preservation solutions in heart transplantation in rats
- Masashi Maeda,
- Naoya Kasahara,
- Junshi Doi,
- Yuki Iijima,
- Takeshi Kikuchi,
- Takumi Teratani,
- Eiji Kobayashi
- Division of Development of Advanced Therapy, Center for Development of Advanced Medical Technology, Jichi Medical University, Tochigi, Japan
- Correspondence to Dr Takumi Teratani, Division of Development Advanced Therapy, Center for Development of Advanced Medical Technology, Jichi Medical University, 3311-1, Yakushiji, Shimotsukeshi, Tochigi 329-0498, Japan;
- Received 1 August 2012
- Revised 29 November 2012
- Accepted 9 December 2012
Objective We developed a novel luciferase-based viability assay for assessing the viability of hearts preserved in different solutions. We examined whether this in vitro system could predict heart damage and survival after transplantation in rats.
Design By our novel system, preserved heart viability evaluation and transplanted heart-graft functional research study.
Setting University basic science laboratory.
Interventions Isolated Luciferase-transgenic Lewis (LEW) rat cardiac-tissue-chips were plated on 96-well tissue-culture plates and incubated in preservation solutions at 4°C. Viability was measured as photon intensity by using a bio-imaging system. Heart-grafts preserved in University of Wisconsin (UW), extracellular-trehalose-Kyoto (ETK), Euro-Collins (EC), histidin-tryptophan-ketoglutarat solution (HTK), lactated Ringer's (LR) or normal saline solution were transplanted cervically by using a cuff-technique or into the abdomens of syngeneic wild-type LEW rats by using conventional microsurgical suture techniques.
Main outcome measures Imaging an evaluation of preservation heart-graft and functional analysis.
Results Cardiac-tissue-chips preserved with UW, HTK or ETK solution gave higher luminance than those preserved with EC, LR or normal saline (p<0.03). After 24 h of preservation of hearts in each solution at 4°C, the beating of the isolated hearts was evaluated. The success rate, evaluation of beating, of cervical heart transplants using UW and ETK solution exceeded 70%, but those using other preservation solutions were lower (UW: 100%, ETK: 75%, EC: 42.86%, HTK: 14.29%, normal saline: 0%). Histological analysis of cervical heart-grafts after 3 h preservation by myeloperoxidase (MPO), zona occludens-1(ZO-1), and caspase-3 immunostaining revealed different degrees of preservation damage in all grafts.
Conclusions Our novel assay system is simple and can test multiple solutions. It should therefore be a powerful tool for developing and improving new heart-graft preservation solutions.
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