Published online Jun 14, 2007. doi: 10.3748/wjg.v13.i22.3133
Revised: February 22, 2007
Accepted: March 8, 2007
Published online: June 14, 2007
AIM: To develop a practical method for isolation, purification and culture of hepatic Kupffer cells (KCs) and to observe their suppressive effects on the proliferation of alloreactive T cells.
METHODS: Perfusion in situ in vivo combined with density gradient centrifugation was applied in isolation, purification and culture of hepatic KC. The suppression by KCs on the T cell proliferation in mixed lymphocyte reaction (MLR) was observed.
RESULTS: This method resulted in a satisfactorily high yield of (1.1 ± 0.2) × 107 KCs per liver, (93.5% ± 1.8%) viable cells, over 90% purity and positive for ED-2. After the first 24 h in culture, a great number of KCs which exhibited typical characteristics were observed. Using 3H-TdR incorporation assay, non-irradiated KCs significantly suppressed allo-MLR. The KCs recovered from accepted liver allografts in groups D and E were more effective in suppressing allo-MLR.
CONCLUSION: A standardized procedure for isolation of highly purified rat KCs is proposed and KCs have suppressive effects on the proliferation of alloreactive T cells, especially those derived from accepted liver allografts.