Establishment of Allogeneic Kidney Transplantation Technical System in Banna Miniature Pig Inbred Strain

doi:10.12300/j.issn.1674-5817.2025.033

Abstract

Abstract:

Objective To establish a technical system for allogeneic kidney transplantation surgery in pigs using the Banna miniature pig inbred strain, and to evaluate it through routine blood tests, liver and kidney function tests, thus providing reference data for the preparation of allogeneic kidney transplantation models. Methods A total of 4 cases of allogeneic kidney transplantation surgeries were performed, including 1 case of single kidney transplantation in a healthy pig, 2 cases of kidney transplantation after unilateral nephrectomy, and 1 case of kidney transplantation after bilateral nephrectomy. Before kidney transplantation, cross-matching and complement-dependent cytotoxicity (CDC) tests were used for matching between donor and recipient pigs. After kidney transplantation, peripheral blood samples were regularly collected from pigs for routine blood tests, liver function tests, and kidney function tests, and color Doppler ultrasound technology was used to detect blood supply to the transplanted kidneys. After reaching the experimental endpoint, both kidneys of pig DR1 and the left kidney of pig DR3 were collected and hematoxylin-eosin (HE) staining was performed to evaluate pathological changes in the transplanted kidneys. Results Recipient pigs DR1 and DR3 died at 17 days and 30 days after surgery respectively, while recipient pigs R and DR2 remained in good condition during the 30-day observation period. The results of liver and kidney function test showed that in pig DR1, alanine aminotransferase (ALT) levels increased on postoperative day 1 (>1 000 U/L), peaked on postoperative day 7 (1 300 U/L), and aspartate aminotransferase (AST) levels peaked on postoperative day 1 (>3 000 U/L). On postoperative day 17, ALT and AST levels remained high (ALT, 500 U/L; AST, 700 U/L). In pigs R, DR2, and DR3, ALT and AST levels returned to normal around day 17. Serum creatinine (Crea) levels in pig R remained stable without postoperative increase. Crea levels in pigs DR1 and DR2 showed transient elevation on postoperative day 1, then gradually returned to normal (<100 μmol/L). Crea levels in pig DR3 remained below 500 μmol/L from postoperative days 2-10, but increased between days 11-28, reaching up to 1 500 μmol/L, indicating gradual loss of kidney function. Ultrasound results showed that the preoperative resistive index (RI) of recipient pig R was 0.91. On postoperative day 24, renal cortex and medulla showed abundant blood flow signals with RI value of 0.88, which was close to the pre-transplantation RI value. For pig DR2, the RI value on postoperative day 17 was 0.89, with poor renal cortex blood flow and relatively good renal medulla blood flow. In pig DR1 on postoperative day 17, no blood flow signals were detected in the transplanted kidney. HE staining results showed that the non-transplanted healthy right kidney of pig DR1 had normal structure, while the transplanted left kidney showed blurred glomerular structure and nuclear dissolution, indicating that the left kidney had lost function before removal. In the transplanted left kidney of pig DR3, large numbers of red blood cells and lymphocyte infiltration were observed in glomeruli and renal tubules, indicating possible coagulation dysfunction and rejection reactions after kidney transplantation. Conclusion Banna miniature pig inbred strain is used as experimental animals to perform four cases of allogeneic pig-to-pig kidney transplantation. The physiological parameters of the recipient pig and the function of the transplanted kidney are monitored after surgery using routine blood tests, liver and kidney function tests, color Doppler ultrasound, and pathological examinations. The allogeneic pig-to-pig kidney transplantation technical system established in the study can provide a foundation for clinicians to conduct kidney transplantation surgeries.

Key words: Banna miniature pig, Inbred strain, Allogeneic kidney transplantation, Technical system

CLC Number:

Q95-33

ZHANG Ying,PENG Ziwei,YANG Chang,et al. Establishment of Allogeneic Kidney Transplantation Technical System in Banna Miniature Pig Inbred Strain[J]. Laboratory Animal and Comparative Medicine, 2025, 45(5): 623-633. DOI: 10.12300/j.issn.1674-5817.2025.033.

Figures/Tables 7

Table 1 Experimental animal grouping for kidney trans-plantation surgery

组别

Group

受体猪编号

Recipient pig number

供体猪编号

Donor pig number

健康猪接受单肾移植

Single kidney transplantation in a healthy pig

单肾切除后肾移植

Kidney transplantation after unilateral nephrectomy

DR1

DR2

DR1

双肾切除后肾移植

Kidney transplantation after

bilateral nephrectomy

DR3

Figure 1 Cross-matching test results between donor and recipient pigsNote： A， Cross-matching results between pig R serum and pig D red blood cells； B， Cross-matching results between pig D serum and pig R red blood cells； C， Cross-matching results between pig DR1 serum and pig DR2 red blood cells； D， Cross-matching results between pig DR2 serum and pig DR1 red blood cells. Scale bar = 50 μm.

Figure 2 Results of the CDC assay between donor and recipient pigsNote： A-B， Flow cytometry results and cell death rate for the donor pig C2， C1， D versus recipient pig R in CDC matching test； C-D， Flow cytometry results and cell death rate for donor pig C2， C1， DR2 versus recipient pig DR1 in CDC matching test； E-F， Flow cytometry results and cell death rate for donor pig C2， C1， DR versus recipient pig DR2 in CDC matching test.

Table 2 Allogeneic pig kidney transplantation surgery conditions

供体猪或受体猪编号 Donor/recipient number	供体肾重量/g Donor kidney weight/g	热缺血时间 Warm ischemia time	冷缺血时间 Cold ischemia time	开放循环后开始排尿时间 Time to initiate urination after opening the loop
R	118.9	1 min 5 s	4 h 35 min	2 min
DR1	87.3	2 min 5 s	1 h 15 min	5 min
DR2	115.7	5 min	6 h 23 min	3 min
DR3	100.5	4 min	8 h 18 min	4 min

Figure 3 Routine blood， liver， and kidney function test results of donor and recipient pigs before and after kidney transplantationNote： WBC， White blood cell count； Lym， Lymphocyte count； Neu， Neutrophil count； ALT， Alanine aminotransferase； AST， Aspartate aminotransferase； Crea， Creatinine. Blood collection in pigs is difficult and may cause stress and detection errors， leading to missing data at certain time points.

Figure 4 Renal blood flow monitoring of donor and recipient pigsNote： A， Preoperative ultrasound image of pig R； B， Postoperative ultrasound image of pig R at 24 days； C， Postoperative ultrasound image of pig DR1 at 17 days； D， Postoperative ultrasound image of pig DR2 at 17 days.

Figure 5 Gross observation and HE staining of transplanted kidneyNote： A-C， Gross appearance of the non-transplanted kidney （contralateral kidney） of pig DR1， transplanted kidney of pig DR1， and transplanted kidney of pig DR3， respectively； D-F， Sectional views of the non-transplanted kidney （contralateral kidney） of pig DR1， transplanted kidney of pig DR1， and transplanted kidney of pig DR3， respectively； G， HE staining of the non-transplanted kidney （contralateral kidney） of pig DR1； H， HE staining of the transplanted kidney of pig DR1， with glomeruli showing nuclear dissolution indicated by black boxes； I， HE staining of the transplanted kidney of pig DR3， with lymphocytes indicated by red boxes and red blood cells indicated by black boxes. Scale bar = 50 μm.

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