Hepatocyte isolation from resected benign tissues: Results of a 5-year experience

Table 1 Patient profile and isolated human hepatocyte viability, total cell yield and hepatocyte yield

No.	Sex	Age (yr)	Blood group	Disease	Tissue weight (g)	Warm ischemia time (min)	Hepatocyte (× 105/g)	Viability (%)
1	Male	48	O+	Hemangiomas	15	30	28.00	62
2	Male	52	A+	Hemangiomas	18	25	20.56	73
3	Female	23	A+	Calculus	21	25	12.86	69
4	Male	55	O+	Hemangiomas	26	30	18.08	65
5	Female	45	A+	Hemangiomas	27	25	51.85	76
6	Female	36	B+	Hemangiomas	32	15	65.63	85
7	Male	47	AB+	Calculus	38	10	71.05	92
8	Male	42	A+	Hemangiomas	42	10	54.76	86
9	Male	52	A+	Hemangiomas	20	5	63.30	87
10	Male	63	O+	Hemangiomas	33	25	72.11	93
11	Female	46	O+	Hemangiomas	27	20	43.70	68
12	Male	57	A+	Hemangiomas	30	35	55.21	75
13	Male	44	B+	Calculus	25	15	63.81	84
14	Male	61	A+	Calculus	17	5	25.20	86
15	Female	57	A+	Hemangiomas	32	7	62.50	89
16	Male	27	A+	Hemangiomas	27	15	52.65	76
17	Female	55	B+	Calculus	23	7	32.07	66
18	Male	61	O+	Hemangiomas	31	25	68.80	90
19	Male	37	B+	Hemangiomas	37	21	72.22	92
20	Male	65	B+	Calculus	20	15	17.07	72
21	Male	79	O+	Calculus	26	20	22.51	66
22	Male	47	O+	Hemangiomas	35	5	67.72	89
23	Male	51	B+	Hemangiomas	24	12	59.27	77
24	Male	67	O+	Hemangiomas	30	17	62.25	82

Table 2 Reagents and solutions preparation (prepared for 80 g liver sample)

Resolution	Ingredient	Concentration (g/L)
PB (Perfusion buffer)	Double-distilled water (3000 mL)
	NaCl (27 g)	9
	KCl (1.26 g)	0.42
	NaHCO3 (6.3 g)	2.1
	Glucose (2.7 g)	0.9
	Hepes (14.34 g)	4.78
PBE (Perfusion buffer with EDTA)	PB (1000 mL)
	EDTA (0.37 g)	0.37
PBD (Perfusion buffer with dispase)	PB (500 mL)
	Dispase II (Sigma) (4.2 g)	8.4
PBC (Perfusion buffer with collagenase)	PB (500 mL)
	Collagenase IV (Sigma) (0.25 g)	0.5
	CaCl2•2H2O (0.275 g)	0.55
WB (Washing buffer)	Double-distilled water (2500 mL)
	NaCl (17.5 g)	7
	KCl (1.15 g)	0.46
	CaCl2•2H2O (0.325 g)	0.13
	Hepes (5.95 g)	2.38
	Bovine serum albumin (2.5 g)	1.0
WBD (Washing buffer with DNase)	WB (Washing buffer) (1500 mL)
	MgCl2•6H2O (0.15 g)	0.1
	MgSO4•7H2O (0.15 g)	0.1
	DNase I (Sigma) (0.15 g)	0.1

Table 3 Concentrations of albumin in the cultured hepatocyte suspension

Time	Albumin (g/L)
24 h	0.79 ± 0.31
7 d	1.36 ± 0.42
10 d	1.09 ± 0.21

The concentration of albumin in the William’s E culture medium was about 0.5 g/L.