47 PHYSIOLOGICAL STATUS OF MALE AND FEMALE MINIATURE PIGS CLONED WITH MESENCHYMAL STEM CELLS

Abstract

The information on the physiological health status and the endocrinological parameters of cloned pigs is limited. To address this issue, the present study evaluated the hematological, biochemical, and endocrinological status of adult cloned male and female miniature pigs. Male and female cloned miniature pigs were produced by NT using mesenchymal stem cells (MSCs) derived bone marrow of miniature pig (T-type, PWG Micro-pig^®, PWG Genetics Korea, South Korea). Cloned and age-matched control male and female miniature pigs were maintained under the same conditions in a farm facility, and collected blood samples via jugular venipuncture at the age of 1 year, 3, 6, and 9 months. Complete blood counts of leukocytes, erythrocytes, and thromocytes were performed using automated hematology cell counter (MS9-5V; Melet Schloesing Lab., France). Biochemical analyses were performed using a bench-top dry chemistry analyzer (Vettest 8008 Chemistry Analyzer; IDEXX Lab., UK) by examining creatinine, glucose, blood urea nitrogen, Gamma-glutamyltransferase (γ–GGT), albumin, total bilirubin, total protein (TP), alanine aminotransferase (ALT), aspartate aminotransferase, creatine kinase, cholesterol, and amylase. Plasma growth hormone, insulin, insulin-like growth factor-1 (IGF-1), thyroid, tyroxine, cortisol, aldosterone, progesterone, testosterone, and estrogen concentration were determined by a 7020 automatic analyzer (Hitachi Ltd., Tokyo, Japan). Most parameters related to the hematological and biochemical status of cloned female and male miniature pigs were similar to control animals. However, γ–GGT (67.0 ± 20.8) and ALT (78.7 ± 24.0) levels of cloned male were higher compared to normal range (16 to 30 and 9 to 43 U L^–1, respectively), and significantly (P < 0.05 and P < 0.01, respectively) higher than cloned female (GGT: 38.7 ± 2.9, ALT: 55.0 ± 16.1) and control female and male pigs (GGT: 27.5 ± 4.8 and 23.0 ± 4.4, ALT: 38.5 ± 7.9 and 32.3 ± 8.5). TP (8.2 ± 0.2) and cholesterol (87.33 ± 6.66) levels of cloned female were higher compared to normal range (6.0 to 8.0 g dL^–1 and 18 to 79 mg dL^–1, respectively), and significantly (P < 0.05) higher than cloned male (TP: 7.7 ± 0.4, cholesterol: 85.0 ± 8.2) and control female and male (TP: 7.9 ± 0.4 and 7.1 ± 0.6, cholesterol: 62.8 ± 3.6 and 57.0 ± 14.4). Endocrinological variation of insulin and IGF-1 of cloned female (1.43 ± 0.7 and 226.10 ± 65.0, respectively) were higher than cloned male and control female and male (0.9 ± 0.1 and 174.2 ± 42.2, 0.5 ± 0.3 and 199.9 ± 8.9, 0.5 ± 0.4 and 168.9 ± 21.2, respectively). In summary, despite similarities in hematological and biochemical parameters between cloned male and female miniature pigs and controls, a greater degree of physiological and endocrinological variations were found in cloned pigs. Based on the changes of the parameters related to growth metabolism, cloned male and female miniature pigs may have dysfunction of the liver. Therefore, the variabilities found must be taken into account before considering the cloned pigs for applications in biomedicine and xenotransplantation.