Isolation of Kupffer cells and their suppressive effects on T lymphocyte growth in rat orthotopic liver transplantation.
OBJECTIVE
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.


CONCLUSIONS
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.
Liu
Hua
H
Department of General Surgery, Renji Hospital, Shanghai Jiao Tong University School of Medicine, No. 1630 Dong Fang Road, Shanghai 200127, China.
Cao
Hui
H
Wu
Zhi-Yong
ZY
D000818:Animals; D002450:Cell Communication / Q000502:physiology; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology / Q000502:physiology; D022781:Hepatocytes / Q000166:cytology / Q000502:physiology; D007728:Kupffer Cells / Q000166:cytology* / Q000502:physiology; D016031:Liver Transplantation / Q000473:pathology*; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D013601:T-Lymphocytes / Q000166:cytology* / Q000502:physiology; D023001:Transplantation Tolerance / Q000502:physiology