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1.
Acta cir. bras ; 39: e390624, 2024. ilus
Article in English | LILACS, VETINDEX | ID: biblio-1533361

ABSTRACT

Purpose: This study aimed to compare the degree of maturation and development of fetal pig segmental intestinal tissue with that of spheroids created by in-vitro reaggregation of dissociated fetal intestinal cells after transplantation into immunodeficient mice. Methods: Fetal pig small intestines were transplanted as segmental grafts into the omentum and subrenal capsules of immunodeficient mice or enzymatically treated to generate single cells. Spheroids made by in-vitro reaggregation of these cells were transplanted into the subrenal capsules of immunodeficient mice. The segmental grafts and spheroids were harvested four and eight weeks after transplantation, and the structural maturity and in-vivo development of these specimens were histologically evaluated. Results: The spheroids were engrafted and supplied blood vessels from the host mice, but an intestinal layered structure was not clearly observed, and there was almost no change in size. On the other hand, the segmental grafts formed deep crypts in the mucus membrane, the inner circular layer, and outer longitudinal muscles. The crypts of the transplanted grafts harvested at eight weeks were much deeper, and the smooth muscle layer and the enteric nervous system were more mature than those of grafts harvested at the fourth week, although the intestinal peristaltic wave was not observed. Conclusions: Spheroids created from fetal small intestinal cells could not form layered structures or mature sufficiently. Conversely, segmental tissues structurally matured and developed after in-vivo transplantation and are therefore potential grafts for transplantation.


Subject(s)
Animals , Mice , Swine , Transplantation, Heterologous/veterinary , Fetal Tissue Transplantation/veterinary , Fetal Organ Maturity
2.
Cryo Letters ; 37(1): 47-52, 2016.
Article in English | MEDLINE | ID: mdl-26964025

ABSTRACT

BACKGROUND: Kidney transplantation from deceased or living human donors has been limited by donor availability as opposed to the increasing demand, by the risks of allograft loss rejection and immunosuppressive therapy toxicity and by limitations of organ preservation protocols, which is essential to organise staff and facilities, transport organs, and perform necessary laboratory tests. However, the cryopreservation of composite tissues poses technical challenges beyond those seen in the preservation of single tissue types or organs. OBJECTIVE: The purpose of our study was to establish a protocol for long-term storing of renal primordia, that generates new adult kidneys after transplant into a syngeneic non-immunosuppressed host. MATERIALS AND METHODS: Metanephroi from 16-days-old embryos were microdissected and vitrified following the minimum essential volume method and using Cryotop as a device and VM3 as vitrification solution. After 3 months of storage in liquid nitrogen (-196 degree C), 20 metanephroi were warmed and transplanted using minimally invasive laparoscopic surgery into retroperitoneal fat of 5-month-old immune-competent New Zealand rabbits. In the same way, 22 fresh metanephroi were transplanted. Twenty-one days after transplantation, hosts were euthanized and developed kidneys were recovered and evaluated morphologically and histologically. RESULTS: Significant growth and fully differentiated mature glomeruli and tubule were observed in all kidney graft explants recovered. In total, 5 metanephroi (25.0%) were successfully grown after vitrification. In the same way, 12 metanephroi (54.5%) were successfully grown in the fresh group. CONCLUSION: These encouraging results reported that metanephroi not only survive vitrification, but they vascularized and developed morphologically normal glomeruli after their allotransplantation. These results suggest that it's possible to create a long-term biobank of kidney precursors as an unlimited source of organs for transplantation, and open new therapeutic possibilities for the patients with chronic renal failure.


Subject(s)
Cryopreservation/veterinary , Fetal Tissue Transplantation/veterinary , Kidney Transplantation/veterinary , Kidney/embryology , Organ Preservation/veterinary , Tissue Banks , Animals , Cryopreservation/instrumentation , Cryopreservation/methods , Female , Organ Preservation/instrumentation , Organ Preservation/methods , Rabbits , Vitrification
3.
Lab Anim Sci ; 46(1): 48-55, 1996 Feb.
Article in English | MEDLINE | ID: mdl-8699819

ABSTRACT

We describe the conditions necessary to reconstitute a bovine immune system in SCID mice that supports the generation of antigen-specific serum antibody responses to a hapten-carrier (DNP-KLH) conjugate. Sublethally irradiated SCID mice were engrafted with fetal bovine liver, lymph node, and thymus. Treatment groups were established in which human recombinant interleukin 7, autologous bovine bone marrow, and engraftment site (omentum or abdominal wall) were evaluated for their ability to enhance the reconstitution of the bovine immune system in SCID mice. Engrafted SCID mice were inoculated at approximately 6 and 12 weeks after engraftment with DNP-KLH. Vaccinated mice were evaluated by enzyme-linked immunosorbent assay for serum antibody specific for DNP-KLH and total concentrations of bovine serum immunoglobulin biweekly through 20 weeks after engraftment. The SCID mice with fetal tissues engrafted onto the abdominal wall from two different donors produced microgram quantities of bovine immunoglobulins and had >70% reconstitution rate. Administration of human interleukin 7 and/or fetal bovine bone marrow did not significantly increase the frequency of reconstitution. Subclass analysis of serum from mice receiving the abdominal implants suggested that class-switching occurred to a predominant non-IgM isotype response after the second inoculation. Pooled thymic and splenic cell populations obtained 20 weeks after engraftment from abdominally engrafted groups yielded 38 to 90% bovine cells, as determined by polymerase chain reaction in situ hybridization with bovine satellite DNA-specific primers. These results indicate that an antigen-specific bovine immune response can be generated in heterochimeric SCID-bo mice.


Subject(s)
Antibody Specificity , Fetal Tissue Transplantation/veterinary , Hematopoietic Stem Cell Transplantation/veterinary , Mice, SCID/immunology , Animals , Antibody Formation/drug effects , Base Sequence , Cattle , DNA, Satellite/analysis , Enzyme-Linked Immunosorbent Assay , Female , Fetal Tissue Transplantation/immunology , Haptens/pharmacology , Hemocyanins/pharmacology , Immunoglobulins/blood , Interleukin-7/pharmacology , Lymphoid Tissue/chemistry , Mice , Molecular Sequence Data , Pregnancy , Retrospective Studies
4.
J Vet Med Sci ; 56(6): 1113-8, 1994 Dec.
Article in English | MEDLINE | ID: mdl-7696401

ABSTRACT

The age-related testicular effect on the ovarian primordia was studied by combined transplantation of fetal testes and ovaries in adult male hosts. First, ovarian primordia of 14-day fetal rats were transplanted into a renal subcapsular position of castrated or intact adult male rats. In both the castrated and the intact hosts, most of the ovarian transplants developed normally with only 3 of them having in part seminiferous tubule-like structures in addition to normal ovarian structure. Second, a 14-day ovary was combined with a fetal testis the age of which varied from 13- to 18-day, and the combination was transplanted. In the combination of a 14-day ovary and a 13-day testis, the results varied in such a way that the ovary or the testis alone developed or otherwise, both gonads developed well. In union with 15- to 18-day testes, the ovaries did not develop, although the testes developed well. These results suggest that the 14-day ovarian primordia have a slight reactiveness to androgens of host rats and that the 13-day fetal testes begin to inhibit the development of the 14-day ovaries co-transplanted with them.


Subject(s)
Fetal Tissue Transplantation/veterinary , Ovary/embryology , Testis/embryology , Animals , Female , Fetal Tissue Transplantation/physiology , Gestational Age , Male , Orchiectomy , Ovary/transplantation , Rats , Rats, Wistar , Testis/physiology , Testis/transplantation
5.
Vet Immunol Immunopathol ; 34(3-4): 273-89, 1992 Nov.
Article in English | MEDLINE | ID: mdl-1455684

ABSTRACT

To develop a model of bovine thymus and lymph node growth in vivo, we have implanted bovine foetal tissues (16-23 weeks gestation) under the renal capsule of severe combined immune deficient (SCID)/beige (BG) mice and assayed for graft growth and characteristics 2-18 weeks after engraftment. Bovine foetal thymus and lymph node grew considerably following engraftment of SCID/BG mice. Growth was optimal if bovine foetal tissues were used before gestation Week 17. Bovine-mouse chimerism was confirmed using glucose phosphate isomerase analysis. Bovine thymus grew during the entire 18 weeks of study. Growth of bovine lymph node was initially rapid, reaching a maximum at 2 weeks after transplantation followed by a progressive decrease in size. Transplanted bovine lymph node and thymus were morphologically similar to age-matched bovine foetal tissue for a limited time period. Fibrosis, degeneration and depletion of lymphocytes were evident 6 weeks after engraftment; changes were more severe in lymph node than in thymus whereas increases in lymphocytes, lymphopoiesis and follicle formation were evident in age-matched bovine foetal tissue. Despite growth and morphological similarities of the transplanted tissue, blood counts suggested there was no peripheralization of bovine leucocytes. Bovine immunoglobulins (IgG1 and IgG2) were detected in serum of some SCID/BG chimeric mice for a limited time. The appearance of bovine immunoglobulins at 2 weeks in SCID/BG chimeric mice depended on the age of the foetal donor (> 18 weeks) and coincided with the appearance of morphologically mature lymphocytes in the donor foetus lymph nodes. The ability to produce bovine immunoglobulins decreased 8 weeks after engraftment, coinciding with the depletion of lymphocytes in the engrafted lymph node. Lymphocyte depletion and loss of function of engrafted tissues appear the result of a lack of lymphoid progenitors normally derived from hematopoietic stem cells in the bone marrow.


Subject(s)
Fetal Tissue Transplantation/veterinary , Lymph Nodes/transplantation , Mice, SCID/immunology , Thymus Gland/transplantation , Animals , Cattle , Chimera/immunology , Fetal Tissue Transplantation/immunology , Graft Survival , Immunoglobulin G/immunology , Kidney/immunology , Leukocyte Count/veterinary , Lymph Nodes/embryology , Lymph Nodes/immunology , Lymphocytes/immunology , Male , Mice , Mice, Inbred BALB C/immunology , Mice, SCID/surgery , Thymus Gland/embryology , Thymus Gland/immunology
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