Foetal hepatocyte transplantation in a vascularized AV-Loop transplantation model in the rat.
The use of foetal liver cells (FLC) in the context of hepatic tissue engineering might permit efficient in vitro expansion and cryopreservation in a cell bank. A prerequisite for successful application of bioartificial liver tissue is sufficient initial vascularization. In this study, we evaluated the transplantation of fibrin gel-immobilized FLC in a vascularized arterio-veno-venous (AV)-loop model. FLC were isolated from embryonic/foetal (ED 16) rat livers and were enriched by using magnetic cell sorting (MACS). After cryopreservation, FLC were labelled by pkh-26. Cells were transplanted in a fibrin matrix into a subcutaneous chamber containing a microsurgically created AV-loop in the femoral region of the recipient rat. The chambers were explanted after 14 days. Subcutaneous implants without an AV-loop and cell-free implants served as controls. Fluorescence microscopy of the constructs was used to identify pkh-26(+)- donor cells. Characterization was performed by RT-PCR and immunhistology (IH) for CK-18 and CD31. Transplantation of FLC using the AV-loop permitted a neo-tissue formation in the fibrin matrix. A high-density vascularization was observed in the AV-loop constructs as shown by CD31 IH. Viable foetal donor cells were detected which expressed CK-18. FLC can be successfully used for heterotopic transplantation. Fibrin matrix permits rapid blood vessel ingrowth from the AV-loop and supports engraftment of FLC. It is therefore an appropriate environment for hepatocyte transplantation in combination with microsurgical vascularization strategies. Transplantation of fibrin gel-immobilized FLC may be a promising approach for the development of highly vascularized in vivo tissue-engineering-based liver support systems.
Fiegel
H C
HC
Department of Pediatric Surgery, University of Leipzig, Germany. henning.fiegel@medizin.uni-leipzig.de
Pryymachuk
G
G
Rath
S
S
Bleiziffer
O
O
Beier
J P
JP
Bruns
H
H
Kluth
D
D
Metzger
R
R
Horch
R E
RE
Till
H
H
Kneser
U
U
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D005260:Female; D005333:Fetus / Q000166:cytology*; D005337:Fibrin / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology / Q000502:physiology / Q000637:transplantation*; D006801:Humans; D018189:Immunomagnetic Separation; D019164:Liver, Artificial; D011247:Pregnancy; D051381:Rats; D011917:Rats, Inbred Lew; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds / Q000737:chemistry
D005337:Fibrin