title: Establishment of major histocompatibility complex homozygous gnotobiotic miniature swine colony for xenotransplantation.
abstract: To overcome shortages of human donor organs for organ failure patients, we made a commitment to develop gnotobiotic miniature swine as an alternative organ donor source for xenotransplantation. For this, we have constructed an absolute barrier-sustained gnotobiotic facility. Pregnant sows of gnotobiotic miniature swine, were procured and germfree piglets were obtained by hysterectomy. These were maintained in germfree isolators for about 4 weeks, deprived of colostrum and were fed sterilized soybean milk. They were associated with di-flora, anaerobic Lactobacillus sp. and Streptococcus sp. After confirmation of successful associations, gnotobiotic piglets were transferred into the facility aseptically. The piglets are maintained on high-efficiency particle air-filtered air in and out; maintaining constant room air pressure of 33 ± 3 mmAq, and sterile water and diet. In 10 sessions of hysterectomy, 18 male and 32 female piglets were obtained of which piglets (M six, F eight) died within 5 days. Among live piglets, piglets (M eight, F 12) were confirmed to be germfree by microbiological monitoring. For research of xenotransplantation, one consistent experimental result was essential. Therefore, major histocompatibility complex class II which related innate immunity, homozygotic gnotobiotic miniature swine was developed. As a result, genotyping revealed 14 individuals to be homozygous for major histocompatibility complex class II (DRB, DQB) as 0301, three individuals were homozygous as 0201 and each of two were homozygous for DQB as 0701 and DRB as 0404, respectively. Genetic modifications and immunological research for ideal alternative organ sources are in progress.
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University, Seoul, Korea; Department of Animal Biotechnology, Konkuk University, Seoul, Korea.
authors.forename: Jeong Ho
authors.initials: JH
authors.lastname: Hwang
authors.forename: Mukesh Kumar
authors.initials: MK
authors.lastname: Gupta
authors.forename: Chan Kyu
authors.initials: CK
authors.lastname: Park
authors.forename: Yoon Berm
authors.initials: YB
authors.lastname: Kim
authors.forename: Hoon Taek
authors.initials: HT
authors.lastname: Lee
mesh_terms: D000818:Animals; D005260:Female; D005856:Germ-Free Life; D006720:Homozygote; D006801:Humans; D008285:Major Histocompatibility Complex / Q000235:genetics*; D011247:Pregnancy; D013552:Swine; D013556:Swine, Miniature / Q000235:genetics*; D014183:Transplantation, Heterologous
keywords: MHC class II homozygous; germ-free piglet; gnotobiotic facility; gnotobiotic miniature swine; xenotransplantation