Treatment of enzyme deficiency by hepatocyte transplantation in rats

Vroemen, J.P.A.M.; Blanckaert, Norbert; Buurman, Wim A.; Heirwegh, Kpm; Kootstra, G.

doi:10.1016/0022-4804(85)90152-0

Cited by 46 publications

(17 citation statements)

References 11 publications

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“…Transplantation of hepatocytes by intrasplenic injection in Gunn rats has been shown to decrease serum bilirubin concentrations (8,23). However, bilirubin levels also decreased after intrasplenic injection of Gunn rat hepatocytes, suggesting that decrease in bilirubin levels might have been due to interference with splenic bilirubin production (9). In a recent study, only minor amounts of conjugated bilirubin were found in the bile of Gunn rats after intrasplenic injection of normal hepatocytes; no bilirubin conjugates were present in bile beyond day 6 after hepatocyte transplantation (10).…”

Section: Resultsmentioning

confidence: 99%

“…Because homozygous Gunn rats lack UDP-glucuronosyltransferase activity for bilirubin, they cannot efficiently excrete bilirubin and, thus, exhibit lifelong nonhemolytic unconjugated hyperbilirubinemia (6)(7)(8). Hepatocyte transplantation has been used to impart the transferase activity in Gunn rats (9,10). However, up to now, long-term function of transplanted hepatocytes was not demonstrated unequivocally.…”

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confidence: 99%

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Survival, organization, and function of microcarrier-attached hepatocytes transplanted in rats.

Demetriou

Levenson

Novikoff

et al. 1986

Proc. Natl. Acad. Sci. U.S.A.

134

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Hepatocytes harvested by collagenase perfusion of rat liver were attached to collagen-coated microcarriers and injected intraperitoneally into congeneic or allogeneic bilirubin-UDP-glucuronosyltransferase (EC 2.4.1.17)-deficient (Gunn) rats or allogeneic analbuminemic (NAR) rats. Five days later, the microcarriers were observed to have formed conglomerates chiefly on the anterior surface of the pancreas. Scanning electron microscopy showed hepatocytes attached to the granular collagen-coated surface of the microcarriers and newly formed connective tissue. Light microscopy revealed that the microcarriers formed a lattice with the collagen tissue; hepatocytes were seen within this lattice or on the surface of the microcarriers. Hepatocyte plasma membranes were nucleoside-diphosphatase (NDPase)-positive. Newly formed blood islands, blood vessels containing erythrocytes and leukocytes and NDPase-positive endothelium were observed in close proximity to the hepatocytes and fibroblasts. Transmission electron microscopic examination showed hepatocytes with microvilli and nucleoid-containing peroxisomes with catalase activity. Hepatocytes were present for up to 2 months in congeneic recipients, the longest period of observation after transplantation. After normal microcarrierattached hepatocytes were transplanted into allogeneic Gunn rats, bilirubin glucuronides were present in bile for 6 days. When congeneic Gunn rat recipients were used, bilirubin glucuronides were present in bile throughout the study (28 days); this was accompanied by reduction of serum bilirubin concentrations to nearly normal levels. After injection of normal hepatocytes into allogeneic NAR rats, plasma albumin concentration progressively increased for 6 days and then declined. In NAR recipients which were immunosuppressed with cyclosporin A, peak plasma albumin levels were reached in 14 days and persisted nearly at that level throughout the study (28 days). We describe a technique in which isolated rat hepatocytes, attached to collagen-coated dextran microcarriers, were injected i.p. in homozygous Gunn rats and genetically analbuminemic (NAR) rats. The injected microcarrierattached hepatocytes form conglomerates in the peritoneal cavity. Light and electron microscopic examination of the conglomerates showed differentiated hepatocytes, newly formed blood vessels, and fibroblasts. Function of the transplanted hepatocytes was shown in Gunn rats by biliary excretion of conjugated bilirubin and reduction of serum bilirubin concentration and in NAR rats by the appearance of albumin in the plasma. MATERIALS AND METHODSMale Wistar rats (200-250 g) were purchased from Charles River Breeding Laboratories. Syngeneic Wistar (RHA) rats and congeneic Gunn rats, which have identical genetic make-up with the Wistar (RHA) rats except for the bilirubin conjugation locus, were developed by C. Hansen of the National Institutes of Health and maintained as congeneic strains at the Albert Einstein College of Medicine (Bronx, NY). Analbuminemic (NAR) rats, mutants...

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Survival, organization, and function of microcarrier-attached hepatocytes transplanted in rats.

Demetriou

Levenson

Novikoff

et al. 1986

Proc. Natl. Acad. Sci. U.S.A.

134

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“…It was shown that liver cell rearrangement after HTX resulted in partial reconfiguration of the hepatic microstructure, with formation of in tercellular bile canaliculi and sinusoids [8,18]. Ample proof was provided that trans planted hepatocytes retain many, and possi bly all, of their metabolic functions [3,4,8,17,18,[27][28][29], In previous work it was shown that in the absence of an excretory route, bile products from transplanted isolated hepato cytes were secreted into the portal circulation, and were eventually excreted by the host liver into the biliary system [3,4,[27][28][29],…”

Section: Introductionmentioning

confidence: 99%

Transplantation of Isolated Hepatocytes into the Pancreas

Vroemen

Buurman²,

Linden³

et al. 1988

Eur Surg Res

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In previous research into hepatocyte transplantation (HTX) the spleen was the preferred acceptor organ for isolated donor hepatocytes. In this study the pancreas was tested as an acceptor organ for HTX. HTX into the pancreas or spleen was performed by injection of 10⁷ isolated hepatocytes into the parenchyma of these organs. Intrapancreatic hepatocytes showed good viability 3 months after syngenic HTX as assessed by histological and immunocytochemical parameters. Definite proof of sustained metabolic activity of normal hepatocytes, 3 months after transplantation into the pancreas of congenitally jaundiced rats, was obtained by demonstration of bilirubin conjugates in bile of the recipients: 4.0% of total biliary bilirubin was conjugated. Intrasplenic HTX, however, was more effective and resulted in a conjugated fraction of 17.7% of total biliary bilirubin (p < 0.001). Reduction of total plasma bilirubin was significant with both methods, but more pronounced in intrasplenic HTX. Bile drainage from the hepatocellular transplant via the pancreatic excretory system into the gut was not observed: conjugated bilirubins were not found in pancreatic juice of HTX-treated jaundiced rats. Intrapancreatic HTX did not adversely affect the host rat; evidence of pancreatitis or diabetes was not found. It is concluded that the pancreas is a suitable acceptor organ for HTX. However, intrapancreatic HTX appears to be less effective than intrasplenic HTX in the treatment of enzyme deficiency disease.

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“…In recent years, the congenitally jaundiced rat has received increased attention from investigators interested in the treatment of Leyten, Vroemen, Blanckaert & Heirwegh inborn errors of metabolism. Transplantation of hepatic tissue from non-jaundiced rats into the GUNN rat appeared to lower plasma bilirubin levels (Rugstad et al, 1970;Matas et al, 1976;Groth et al, 1977;Sutherland et al, 1977;Vroemen et at., 1985a). In many studies in which the GUNN rat was used, the reproducibility of results was poor because of the genetic variability of the existing GUNN rat strains and the lack of a non-deficient but otherwise identical control rat strain.…”

Section: Discussionmentioning

confidence: 99%

The congenic normal R/APfd and jaundiced R/APfd-j/j rat strains: a new animal model of hereditary non-haemolytic unconjugated hyperbilirubinaemia due to defective bilirubin conjugation

et al. 1986

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In this paper the production of the R/APfd-j/j strain which is congenic with the R/APfd strain is reported. The R/APfd-j/j completely lacks hepatic bilirubin UDP-glucuronyltransferase activity, as do our GUNNXR/Pfd-j/j rat strain and various other stocks of GUNN rats (j/j) described in the literature. Our recombinant inbred strain GUNNXR/Pfd-j/j was produced from non-inbred GUNN (j/j) rats. This GUNNXR/Pfd-j/j rat was used as a donor of the jaundice gene j, the R/APfd rat serving as the recipient. After eight backcross-intercross cycles (16 generations) the R/APfd-j/j strain was obtained which is congenic with the R/APfd strain. Congenicity was demonstrated by various techniques including transplantation of skin tissue, strain-specific tumour cells and hepatocytes, the mixed lymphocyte reaction, and comparison of biochemical markers. The potential of the novel inbred strain of jaundiced rat, R/APfd-j/j, and the corresponding control strain R/APfd for biochemical and clinical studies of bilirubin metabolism are briefly discussed.

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Treatment of enzyme deficiency by hepatocyte transplantation in rats

Cited by 46 publications

References 11 publications

Survival, organization, and function of microcarrier-attached hepatocytes transplanted in rats.

Survival, organization, and function of microcarrier-attached hepatocytes transplanted in rats.

Transplantation of Isolated Hepatocytes into the Pancreas

The congenic normal R/APfd and jaundiced R/APfd-j/j rat strains: a new animal model of hereditary non-haemolytic unconjugated hyperbilirubinaemia due to defective bilirubin conjugation

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