Optimization of the technique to isolate fetal hepatocytes, and assessment of their functionality by transplantation.
In contrast to adult hepatocytes, fetal hepatocytes (FH) are thought to be highly proliferative less immunogenic and more resistant to both cryopreservation and ischemic injury. In the present study, we describe the method for isolation of FH and the relationship between the transplantability of FH into the spleen of analbuminemic rats and expression of albumin mRNA. Rat FH were obtained using the nonperfusion collagenase/DNase digestion method. Nagase analbuminemic rats (NAR), a strain which bears a mutation that determines the impossibility of the normal splicing of the albumin mRNA were used as recipients. The transplanted FH immediately migrated to the liver via portal vein, and anchored there. To assess the functional state of the transplanted cells, one month after transplantation, the expression of the albumin gene was studied in the liver of the recipients.
Arahuetes
R M
RM
Departamento de Biología Animal II, Facultad de Biología, Universidad Complutense, Madrid, Spain. rportero@eucmax.sim.ucm.es
Sierra
E
E
Codesal
J
J
Garcia Barrutia
M S
MS
Arza
E
E
Cubero
J
J
Ortiz
A
A
Maganto
P
P
D000255:Adenosine Triphosphate / Q000378:metabolism; D000418:Albumins / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D015152:Blotting, Northern; D002469:Cell Separation; D017690:Cell Transplantation / Q000502:physiology*; D005260:Female; D016332:Fetal Tissue Transplantation / Q000502:physiology*; D005456:Fluorescent Dyes; D022781:Hepatocytes / Q000637:transplantation*; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D011247:Pregnancy; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017208:Rats, Wistar
D000418:Albumins; D005456:Fluorescent Dyes; D012333:RNA, Messenger; D000255:Adenosine Triphosphate