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ISOLATION AND CULTURE OF SINUSOIDAL CELLS FROM RAT AND HUMAN LIVER NEEDLE BIOP- SIES H. Van Bossuyt, M. Reekmans, P. Van der Spek X, E. Wisse Lab. Cell Biology and Histology, Free University of Brussels (VUB), XDepartment of Internal Medicine, Academic Hospital, Laarbeeklaan, 1090 Brussels-Jette, Belgium.

The role of sinusoidal cells in the pathogenesis of l i ver diseases is almost unknown. Iso- lation of these cells might contribute to a better understanding of their role. Sinusoidal cells have been isolated from whole human l ivers. Due to the low ava i lab i l i ty of donor organs, we worked out a method for the isolation and culture of sinusoidal cells from rat and human l iver needle biopsies.

Needle biopsies are puncture perfused with collagenase and pronase in GBSS. Tissue blocks are further incubated in this medium. The resulting cell suspension is purified by centrifu- gation on a Metrizamide density gradient and cultured in Dulbecco's MEM. Using rat l i ve r needle biopsies, different types of sinusoidal cells can be recognized in TEM or SEM after 20 h of culture : endothelial cells are vacuolated and show fenestrae, Kupffer cells have an irregular shape and many dense bodies, are positive for peroxidase staining and avidly phago- cytose latex particles, fat storing cells are recognized by their fat droplets and RER, and pi t cells can be recognized by their specific granules. Using human l i ve r needle biopsies, we have so far only been able to isolate Kupffer cells, which showed the same morphology as rat Kupffer cells.

The usefulness of puncture perfusion was clearly demonstrated by a higher yield of well spread cells (x 1.5). The yield was calculated to vary between 1 and 4.105/g l i ver .

In this study, we were able to demonstrate that sinusoidal cells can be isolated from l iver needle biopsies, by applying a puncture perfusion technique. Four different types of cells could be recognized in co-culture, using morphological, functional or cytochemical cr i ter ia . This technique might be of great help in the study of sinusoidal cells in the pathogenesis of l i ver diseases, such as the interaction with viruses or endotoxin.

ALPHA-I-ANTITRYPSINE (AAT) PHENOTYPE CONVERTS IMMEDIATELY AFTER ORTHOTOPIC LIVER TRANSPLANTATION (OLT).

328 A.B.M.M.v.d. Putten, J.A. Kramps**, J.R. Huizensa , R.A.F. Krom*, R. v. Furth**~ C.H. Gips. Depts. of Medicine and Surgery*, University Hospital Croningen, Depts of Infectious Diseases*** and Pulmonology**, University Hospital Leiden (The Netherlands).

AAT-deficiency can constitute an indication for OLT and conversions of phenotype in the recipient has been reported (NEJM 1980, 302, 272-275). In an adult consecutive OLT-series we have studied these phenomenes systematically. Sere for phenotyping were collected in donor and recipient at donorannouncement and i, 2, 3, 7, 14, 28 days and 3 months after OLT. Of 37 OLT recipients one with AAT-deficiency had type Z, 31 M, 4 MS, 1MI. Of 37 donors one is unknown, 30 M, 2 MZ, 4 MS. Two re-OLT recipients had M and their donors had M too. The median serum AAT of the recipients was 295 g/e and of the donors 260 g/e. After exclusion of 26 M combinations in OLT and re-OLT there were ii chances for detectable conversions of AAT phenotype after OLT. Conversion occurred after 3 days for 3 of the 4 MS recipients, for the other 8 patients conversion was already seen one day after OLT.

Conclusions: The AAT-phenotype invariably changes after OLT to that of the donor and the overruling importance of the donor liver in this process is expressed by the extremely rapid conversion after OLT.

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