Small animal ex vivo machine perfusion of the liver: A comprehensive literature review

Table 4 Perfusate compositions in small animal liver ex vivo machine perfusion studies

Ref.	Animal weight	Liver weight	Perfusate volume	Perfusate composition
Kim et al[14]	200-400 g	NA	200 mL	Cold UW-G solution
Dutkowski et al[15]	250-300 g	10.4 ± 0.3 g	500 mL	Modified UW solution: Starch and glutathione was omitted, supplemented with 80 mg/L gentamycin, and 5000 IU/L heparin
Compagnon et al[67]	300 ± 50 g	NA	150 mL	HMP: Celsior-HES solution. NMP: KHB buffer with 5% bovine serum albumin
Lauschke et al[16]	250-300 g	NA	125 mL	HMP: HTK or Belzer MPS solution supplemented with 6000 IU superoxide dismutase. NMP: KH buffer
Lee et al[90]	200-250 g	NA	100 mL	KH solution, then switched to UW solution (starch omitted)
Tan et al[91]	180-220 g	NA	120 mL	Modified Hoffmann perfusate: Hydroxyethyl starch 50 g/L, calcium gluconate 80 mmol/L, raffinose 10 mmol/L, KH2PO4 25 mmol/L, hydroxyethyl piperazine 10 mmol/L, dexamethasone 12 mg/L, penicillin 2 × 105 units/L, insulin 100 units/L, and with/without 25 mmol/L MgCl and/or 5 mmol/L ATP
Xu et al[92]	200-250 g	NA	NA	HMP: UW solution with/without starch. NMP: KH buffer solution containing 112 μmol/L taurocholic acid, and 150 μg/L hyaluronic acid
Bessems et al[17]	350 ± 50 g	16.5 ± 0.5 g	250 mL	HMP: The UW-G solution or polysol. NMP: KHB without bovine serum albumin
Dutkowski et al[18]	250-300 g	10.8 ± 1.4 g	450 mL	HOPE: Modified starch-free UW solution. NMP: KHB buffer
Tolboom et al[19]	250-300 g	9.74-0.81 g	55-60 mL	NMP: Phenol red-free WEM supplemented with 2 IU/L insulin, 40000 IU/L penicillin, 40000 mg/L streptomycin, 0.292 g/L L-glutamine, 10 mg/L hydrocortisone, 1000 IU/L heparin with 25% (v/v) freshly isolated rat plasma and freshly isolated rat erythrocytes to a hematocrit of 16% to 18%
Vairetti et al[20]	250-300 g	NA	200 mL	KH solution with 1.25 mmol/L CaCl2 or with 0.25 mmol/L CaCl2
Manekeller et al[21]	250-300 g	NA	125 mL	HMP: HTK or Blezer MPS solutions. NMP: KH buffer
Stegemann et al[22]	250-300 g	NA	HMP: 125 mL; NMP: 250 mL	HMP: HTK or modified HTK solution (custodiol-N). NMP: KH buffer with 3 g/100 mL of bovine serum albumin
Ferrigno et al[23]	375 ± 15 g and 300 ± 10 g	NA	200 mL	KH medium
Lüer et al[24]	250-300 g	NA	125 mL	HMP: HTK solution. NMP: WEM solution, supplemented with 3 mg/100 mL of bovine serum albumin
Olschewski et al[25]	250-280 g	NA	NA	HMP, SNMP: Lifor solution. NMP: KH solution
Minor et al[26]	250-300 g	NA	HMP: 125 mL; NMP: 250 mL	HMP: HTK solution supplemented with 0 μmol/L, 10 μmol/L, 50 μmol/L or 100 μmol/L of dopamine. NMP: WEM solution with 3 mg/100 mL of bovine serum albumin
Tolboom et al[27]	250-300 g	NA	55-60 mL	WEM with autologous erythrocytes and plasma. To this were added: Insulin (2 IU/L), penicillin (40000 IU/L)/streptomycin (40000 μg/L), L-glutamine (0.292 g/L), hydrocortisone (10 mg/L), and heparin (1000 IU/L)
Schlegel et al[30]	250-300 g	9.83 ± 0.95 g	50 mL	Modified starch free UW-solution
Giannone et al[28]	250-300 g	NA	NA	Celsior solution
Perk et al[29]	200-300 g	NA	55-60 mL	Phenol red-free WEM supplemented with 2 IU/L insulin (28.85 units/mg), 100000 IU/L penicillin, 100 mg/L streptomycin sulfate, 0.292 g/L L-glutamine, 10 mg/L hydrocortisone, and 1000 IU/L heparin. Fresh rat plasma (25% v/v) and erythrocytes (18%-20% v/v) were collected and added to the perfusate
Bruinsma et al[88]	180-250 g	NA	500 mL	WE supplemented with insulin (2 IU/L), penicillin (40000 IU/L)/streptomycin (40000 μg/L), L-glutamine (0.292 g/L), hydrocortisone (10 mg/L)
Liu et al[31]	NA	21.4 ± 3.6 g (control) 22.9 ± 5.8 g (defatting group)	200 mL	The control perfusate: Minimum essential medium supplemented with 3% wt/vol bovine serum albumin, 1.07 mmol/L lactic acid, and 0.11 mmol/L pyruvic acid. The defatting perfusate: The control perfusate supplemented with the 6 defatting agents (forskolin, GW7647, scoparone, hypericin, visfatin, and GW501516)
Carnevale et al[93]	250-300 g	NA	250 mL	HMP: HTK solution: 100 mmol/L sodium gluconate, 7 mmol/L potassium gluconate, 20 mmol/L sucrose, 30 mmol/L BES, 2.5 mmol/L KH2PO4, and 0.03 mmol/L polyethylene glycol (35 kDa), 5 mmol/L MgSO4, 3 mmol/L glutathione, 5 mmol/L adenosine, and 15 mmol/L glycine, together with 0.25 mg/mL streptomycin and 10 IU/mL penicillin G. NMP: KH buffer with 4% dextran added
Schlegel et al[32]	250-320 g	9.7 ± 1.5 g	50 mL	Modified starch free UW-solution
Schlegel et al[33]	250-320 g	10.14 ± 2.73 g	50 mL	NMP: Diluted full blood or leukocyte and platelet depleted blood perfusate. HOPE: Modified starch-free UW solution
Bae et al[34]	300 ± 25 g	NA	100 mL	HMP: KPS-1 solution (Identical to Belzer’s UW MPS) or KPS-1, enhanced with α-ketoglutarate, L-arginine, N-acetylcysteine, nitroglycerin, and prostaglandin E1 or KPS-1, with 5.4 × 10-2 mmol/L of α-tocopherol diluted in acetone. NMP: KHB buffer
Niu et al[68]	261 ± 4 g	NA	353 ± 11 mL	KH buffer
Tarantola et al[35]	375 ± 15 g and 300 ± 10 g	NA	200 mL	KH medium
Bruinsma et al[36]	250-300 g	Approximately 10 g	100 mL	3-OMG loading solution: 500 mL phenol-red free WEM, 5 mL of 200 mmol/L L-glutamine (0.292 mg/L), 4 mL of penicillin-streptomycin (5000 IU/mL), 5 mg of hydrocortisone, 5000 U of sodium heparin, 375 U of insulin, 19.42 g of 3-O-methyl glucose (0.2 M)
Ferrigno et al[37]	250-300 g	NA	NA	A modified KH buffer
Jia et al[38]	250-300 g	NA	60 mL	UW or HTK solutions
Westerkamp et al[94]	290-320 g	NA	100 mL	HMP, SNMP, rewarming MP: Belzer MPS. NMP: 25 mL of human red blood cell concentrate (final hematocrit 25%), 53.9 mL of WEM solution, 20 mL of human albumin (200 g/L), 1 mL of insulin (100 IU/mL), and 0.1 mL of unfractionated heparin (5000 IU/mL)
Carbonell et al[95]	225-250 g	NA	NA	KH buffer: 118 mmol/L NaCl, 4.7 mmol/L KCl, 1.2 mmol/L MgSO4, 1.2 mmol/L KH2PO4, 2.5 mmol/L CaCl2, 25 mmol/L NaHCO3, 20 mmol/L HEPES
Op den Dries et al[75]	303 ± 4 g (mean ± SEM)		100 mL	20 mL of human red blood cell concentrate (final hematocrit 15%-20%), 59 mL of WEM solution, 20 mL of human albumin (200 g/L), 1 mL of insulin (100 IU/mL), and 0.1 mL of unfractionated heparin (5000 IU/mL)
Okamura et al[39]	250-300 g	NA	300 mL	SNMP: Polysol solution. NMP: KH buffer
Berardo et al[40]	250-300 g	NA	NA	KH buffer
Chai et al[89]	250-300 g and 600-630 g	NA	80 mL	UW solution with/without 0.165 mg/L of metformin
Zeng et al[41]	250-300 g	NA	150 mL	HTK solution
Beal et al[42]	250-350 g	NA	300 mL	86 mL of 25% albumin, 184 mL of WEM, 30 mL of penicillin/streptomycin (10 IU/mL penicillin and 0.01 mg/mL streptomycin), insulin (50 IU/L), heparin (0.01 IU/mL), L-glutamine (0.292 g/L), and hydrocortisone (0.010 g/L). Addition of 625 IU/mL pegylated-catalase
Tabka et al[69]	250-300 g	NA	SNMP: 150 mL; NMP: 140 mL	SNMP: Celsior with/without Ang IV. NMP: KBB enriched with 5% albumin
Xue et al[43]	250 ± 10 g	NA	HMP: 100 mL; NMP: 250 mL	HMP: HTK solution. NMP: KH buffer with 4% dextran
He et al[78]	250-300 g	NA	NA	HTK solution
Gassner et al[70]	280-350 g	13.6 g ± 2.15 g	50 mL	NMP: Low-glucose DMEM supplemented with the rat erythrocyte concentrate and 12.5 mL strain specific rat plasma
Zeng et al[79]	20-24 g	NA	50 mL	HTK solution
Jia et al[80]	250-300 g	NA	60 mL	HTK solution
Oldani et al[44]	176 (155-193) g	NA	50 mL	HOPE: IGL-1 solution with 150 IU heparin. NMP: 25 mL Fischer rat blood with 25 mL normal saline, and with 150 IU heparin
Chin et al[45]	200-250 g	NA	150 mL	DMEM supplemented with 200 mmol/L L-glutamine, 10% v/v FBS, 5% with bovine serum albumin, 8 mg/L dexamethasone, 2000 IU/L heparin, 2 IU/L insulin, and 5 × 106 engineered rat fibroblasts
Gillooly et al[46]	Approximately 300 g	NA	100 mL	99 mL WEM with 10 IU insulin, and lipid nanoparticles (50 nM siRNA)
Martins et al[47]	320-350 g	NA	NA	50% plasma-Lyte and 50% KH solution
Scheuermann et al[48]	296 g ± 8 g (mean ± SEM)	NA	100 mL	SNMP, NMP: 80 mL of WEM supplemented with 5% bovine serum albumin, 20 mL of type O + human RBCs, 0.2 IU insulin, 29.2 mg L-glutamine, 1 mg hydrocortisone, and 500 IU heparin. NMP: KH buffer supplemented with 5% bovine serum albumin
Claussen et al[71]	280-350 g	NA	50 mL	10 mL of the erythrocyte concentrate, 35 mL of DMEM, 5 mL of strain specific rat plasma supplemented with 1000 IU heparin and 12 mmol/L glycine
Haque et al[49]	250-300 g	NA	500 mL	NMP: 950 mL of William’s E media, 20 g of bovine serum albumin, 20 g of polyethylene glycol 35000, 20 mg of dexamethasone, 2 mL of heparin, 1 mL of regular insulin, 10 mL penicillin-streptomycin, 10 mL of antibiotic-antimycotic, and 2.2 g of sodium bicarbonate
Schlegel et al[50]	250-320 g	9.8 0.6 g	100 mL	HOPE: Belzer MPS. NMP: Belzer MPS or diluted heparinized blood
Nösser et al[51]	398.87 ± 133.12 g	14.15 ± 2.66 g	250 mL, 100 mL, 80 mL, 50 mL	SNMP: 500 Ml DMEM supplemented with 100 μg/mL penicillin and streptomycin, 4 mmol/L L-glutamine/L-alanine, 1 μM human insulin, 14 ng/mL glucagon, 1 μM dexamethasone. NMP: The isolated RBCs suspended in DMEM, isolated rat plasma (10% of the total volume), and 500 IU of heparin
Yamada et al[96]	260-350 g	DCD 10.12 ± 0.93 g	NA	KH buffer
Hu et al[52]	250-300 g	NA	150 mL	HTK solution
Von Horn and Minor[53]	250-300 g	NA	200 mL	Aqix RS-I solution
Raigani et al[54]	NA	NA	500 mL	High-glucose DMEM supplemented with 10% v/v FBS, 2% v/v penicillin-streptomycin, and 3% w/v bovine serum albumin. Defatting cocktail agents include 10 μM forskolin, 1 μM GW7647, 1 μM GW501516, 10 μM scoparone, 10 μM hypericin, 0.4 ng/mL visfatin, 0.8 mmol/L L-carnitine, and additional amino acids
Yang et al[55]	200-220 g	NA	NA	60 mL DMEM/F12 (1:1) containing 20% FBS and 1% penicillin-streptomycin solution (penicillin 10000 IU/mL, streptomycin 10000 μg/mL), 20 mL of fresh blood, 5 IU/mL of heparin, 2 IU/L of insulin, and 2.5 μg/mL of dexamethasone
Westerkamp et al[76]	270-300 g	NA	100 mL	25 mL human red blood cell concentrate, 53.9 mL WEM solution, 20 mL human albumin (200 g/L), 1 mL insulin (100 IU/mL), and 0.1 mL unfractionated heparin (5000 IU/mL)
De Vries et al[86]	200-250 g	NA	500 mL	WEM supplemented with sodium bicarbonate (2.2 g/L), dexamethasone (24 mg/L), insulin (5 IU/L), heparin (2000 IU/L), and bovine serum albumin (10 mg/mL)
Liu et al[85]	NA	NA	70 mL	HMP: UW solution with 250 IU heparin, 20000 IU penicillin and 2 mg hydrocortisone. Addition of CIRP competitive inhibitor (C23, 300 ng/mL). NMP: 20 mL rat blood, 50 mL WEM, 250 IU heparin, 20000 IU penicillin, 2 mg hydrocortisone, 0.4 IU insulin, and 0.0292 g glutamine
Lin et al[56]	180 ± 20 g	NA	50 mL	HTK solution with or without various defatting agents (10 mmol/L forskolin; 1 mmol/L GW7647; 10 mmol/L hypericin; 10 mmol/L scoparone; 0.4 ng/mL visfatin; 1 mmol/L GW501516)
Rigo et al[57]	200-250 g	10.35 g (0.41) mean (SEM)	70 mL	NMP: 20 mL of fresh rat blood, 50 mL of complete phenol red-free WEM, supplemented with 11.6 mmol/L glucose, 50 IU/mL penicillin, 50 μg/mL streptomycin, 5 mmol/L L-glutamine, 1 IU/mL insulin, 1 IU/mL heparin, and 2 mEq of sodium bicarbonate
Xu et al[97]	Approximately 500 g	Approximately 15 g	60-110 mL	Whole blood-based perfusate with different defatting components
Carlson et al[58]	331 ± 16 g (mean ± SEM)	NA	95 mL	WEM, 3250 IU each penicillin/streptomycin, 0.65 mmol/L sodium pyruvate, 1.30 mmol/L L-glutamine, 1% human albumin, 500 IU heparin, 15 mg papaverine, 1 mg insulin, 1.25 mg hydrocortisone, and 30 mL of leukoreduced, packed RBCs
Zhou et al[81]	250-300 g	NA	NA	NA
Cao et al[59]	200-220 g	NA	80 mL	DMEM/F12, 20 mL rat blood, 100 IU/mL penicillin, 100 μg/mL streptomycin, and 5 IU/mL heparin
De Stefano et al[60]	200-250 g	Control group: 17.10 (1.93). WI group: 15.17 (0.83)	150 mL	100 mL of phenol red-free WEM, supplemented with 100 IU/mL penicillin, 100 μg/mL streptomycin, 0.292 g/L L-glutamine, 1 IU/mL insulin, 1 IU/mL heparin, and 50 mL of recently expired (max 5 days) human red blood cell
Sun et al[61]	200-220 g	NA	2 mL	Normal saline or single cell suspension containing 1× 107-3 × 107 BMMSCs
Jennings et al[62]	320 ± 11 g (mean ± SEM)	NA	130 mL or 146 mL	WEM (65 mL for pRBC oxygen carriers and 50 mL for oxyglobin) with 3250 U each penicillin/streptomycin, 500 U heparin, 1 mg insulin, 1.25 mg hydrocortisone, and 15 mg papaverine. In addition, sodium pyruvate 0.65 mmol/L, L-glutamine 130 mmol/L, and human albumin 1%. The oxygen carriers added to the perfusate were human pRBC (30 mL), rat pRBC (30 mL), or oxyglobin (46 mL)
Wang et al[72]	NA	NA	HOPE: 50 mL; NMP: 30 mL	HOPE: HTK solution. NMP: 7.5 mL blood, 22.5 mL KHB solution containing 2% FBS, 60 mg glucose, 1 IU insulin, and 150 IU heparin
Asong-Fontem et al[73]	NA	NA	NA	HOPE: IGL-2 or PERF-GEN (Belzer MPS) solutions. NMP: Williams medium E supplemented with insulin 2 U/L, penicillin (40000 U/L)/streptomycin (40000 μg/L), L-glutamine, hydrocortisone 10 mg/L and heparin (1000 U/L)
Shi et al[77]	320-350 g	NA	36 mL	24 mL whole heparinized blood supplemented with 10% sodium citrate, 1% penicillin and streptomycin, and 12 mL circuit priming solution with 45% lactated ringer, 5% sodium bicarbonate and 50% hydroxyethyl starch
Von Horn et al[63]	250-300 g	NA	150 mL	HMP: Aqix RS-I solution. NMP: WEM supplemented with 3 mg/100 mL of bovine serum albumin
Zhou et al[82]	250-300 g	NA	NA	NA
Luo et al[83]	250-300 g	NA	HMP: 150 mL; NMP: 180 mL	HMP: HTK solution. NMP: DMEM/F12, 20% FBS, 1% penicillin streptomycin solution (penicillin 10000 IU/mL, streptomycin 10000 mg/mL), 5 IU/mL of heparin, 2 IU/L of insulin, and 2.5 mg/mL of dexamethasone. Full rat blood (30-45 mL) was reconstituted up to a total volume of 180 mL perfusate
Ohara et al[64]	250-350 g	NA	100 mL	Rat whole blood
Chen et al[65]	34 ± 4 g (mean ± SEM)	Approximately 1 g	300 mL	WEM was supplemented with 20% FBS, 1% penicillin/streptomycin, 5000 IU/L heparin, 50 IU/L insulin, and 0.010 g/L hydrocortisone
Fukai et al[74]	NA	NA	300 mL	HMP: UW-MPS solution. NMP: KHB solution
Hughes et al[84]	250-300 g	14 ± 0.2 g	150 mL	Phenol red-free WEM with addition of 25% bovine albumin, rat RBCs, 2 IU/L insulin, 40000 IU/L penicillin, 40000 μg/L streptomycin, 0.292 g/L L-glutamine, 10 mg/L hydrocortisone, 1000 IU/L heparin, and bicarbonate 75%
Bai et al[98]	360-380 g	NA	36 mL	24 mL whole heparinized blood supplemented with 10% sodium citrate, 1% penicillin, and streptomycin and 12 mL circuit priming solution with 45% lactated ringer, 5% sodium bicarbonate, and 50% hydroxyethyl starch
Von Horn et al[66]	250-300 g	NA	NA	Rewarming MP: Diluted Steen solution or Belzer MPS. NMP: WEM supplemented with 3 mg/100 mL of bovine serum albumin
Li et al[87]	250-350 g	NA	100 mL	90 mL WEM with 10 mL rat blood cells supplemented with 10 mg/L hydrocortisone, 5 IU/mL heparin, 1 IU/mL insulin, 5 mmol/L L-glutamine, 40 IU/mL penicillin, 40 μg/mL streptomycin, and with/without 10 mmol/L epigallocatechin gallate

MP: Machine perfusion; HMP: Hypothermic machine perfusion; NMP: Normothermic machine perfusion; NA: Not written or not applicable in the main article; SNMP: Subnormothermic machine perfusion; DCD: Donated after circulatory death; WI: Warm ischemia; HOPE: Hypothermic oxygenated machine perfusion; UW: University of Wisconsin; UW-G: University of Wisconsin-gluconate; HES: Hydroxyethyl starch; KHB: Krebs Henseleit bicarbonate; HTK: Histidine Tryptophan Ketoglutarate; Belzer MPS: Belzer machine perfusion solution; KH: Krebs Henseleit; KH₂PO₄: Potassium dihydrogen phosphate; MgCl: Magnesium chloride; ATP: Adenosine triphosphate; WEM: William’s E Medium; CaCl₂: Calcium chloride; MgSO₄: Magnesium sulfate; KPS: Kidney perfusion solution; 3-OMG: 3-O-methyl-D-glucose; NaCl: Sodium chloride; KCl: Potassium chloride; NaHCO₃: Sodium bicarbonate; HEPES: 4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid; Ang IV: Angiotensin IV; KBB: Krebs bicarbonate buffer; DMEM: Dulbecco’s modified Eagle’s medium; IGL: Institute George Lopez; siRNA: Small interfering RNA; FBS: Fetal bovine serum; RBCs: Red blood cells; CIRP: Cold-inducible RNA-binding protein; BMMSC: Bone marrow mesenchymal stem cell; pRBC: Packed red blood cell.