Hua Liu1, Hui Cao, Zhi-Yong Wu. 1. Department of General Surgery, Renji Hospital, Shanghai Jiao Tong University School of Medicine, No. 1630 Dong Fang Road, Shanghai 200127, China.
Abstract
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) multiply 10(7) 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 (3)H-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.
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) multiply 10(7) 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 (3)H-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.
Authors: Markus Rentsch; Kerstin Puellmann; Slawo Sirek; Igors Iesalnieks; Klaus Kienle; Thomas Mueller; Ulrich Bolder; Edward Geissler; Karl-Walter Jauch; Alexander Beham Journal: Transpl Int Date: 2005-09 Impact factor: 3.782