Establishment of chronic graft-versus-host disease models after allo-bone marrow transplantation in rats / 中国组织工程研究

ABSTRACT

AIM: To investigate the feasibility of establishment for chronic graft-versus-host disease (cGVHD) model in rats after allo-bone marrow transplantation (allo-BMT), in order to provide premise conditions for further studying the immuno-regulation role of mesenchymal stem cell (MSC) on GVHD after allo-BMT. METHODS: This experiment was finished in Laboratory of Pathology and Pathophysiology in Sun Yat-sen University from April to September 2006.①Six-week-old male Fischer344 rats (RT1Al) were used as donors while six-week-old female Waster (RT1Au) rats were used as recipients.②Recipient rats were given water supplemented with gentamycin (320 mg/L) and erythromycin (250 mg/L) three days before BMT. On the day of transplantation, recipient rats received 8.0 Gy (60Co ?, 0.7 Gy/min) total body irradiation. Within 6 hours following the irradiation, recipient rats in BMT group were transplanted with 0.8?108 cells via tail vein injection, while rats in control group only received the same volume injection of phosphate buffer. Each group included 10 animals. Evaluation of common living status was monitored including daily diet, activity, stool and urine, fur and body mass. Shaved skin, liver and intestine tissues were also analyzed histologically. RESULTS: ①All rats in the control group died within 17 days following the irradiation and most of them died on day 11 or 12 post-transplantation, while BMT group had higher survival rate, in addition to three rats died on days 12, 16, 18 respectively, whereas others were all alive through 60 days expectation period.②Rats in the BMT group had no clear symptoms of acute GVHD, such as rapid weight loss and severe diarrhea, however, the weight growth in rats of the BMT group was quite slow. Furthermore, 1 month following BMT, depilation phenomenon was evident in the head and back of recipients, and then extended to abdominal part and extremity with the increase of time. Two months following BMT, skin follicular dropout and slight dermal mononuclear infiltration were found. Hepatic disease was characterized by portal tract lymphocyte infiltration, fibrous thickening and sclerosis of the bile duct wall. Small bowel specimens showed clear inflammatory cell infiltration (neutrophils, acidophils, macrophages) within lamina propria. CONCLUSION: ①The cGVHD model can be established through allo-BMT from F344 to Wistar rats.②The typical histological signs of cGVHD are evident in skin, liver and intestine tissues, among which hepatic sign is the most dominant including portal tract and bile duct mononuclear infiltration followed by fibrous thickening and sclerosis of the bile duct wall.