Valores hematológicos e bioquímicos de camundongos imunossuprimidos BALB/c NUDE E C57BL/6 SCID / Haematological and biochemical values of immunosuppressed BALB/c NUDE and C57BL/6 SCID mice

Background: The emergence of the NUDE and SCID immunosuppressed mice lineages generated knowledge on various mechanisms of lymphocyte maturation and human autoimmune diseases. Information on haematological and biochemical parameters of these lineages is still scarce, making it impossible to infer homeostasis by comparing data, or to detect genetic influences on the parameters for these species. Haematological and biochemical tests were carried out on Balb/c NUDE and C57BL/6 SCID mice of both sexes, aiming to analyse the presence of genetic influence on possible variations of such parameters and to verify reference values for both lineages.Materials, Methods & Results: One hundred and forty mice (Mus musculus) of the Balb/C NUDE and C57BL/6 SCID lineages were used in the present study. The animals were previously anesthetized, the blood collection procedure was performed by cardiac puncture and the samples were collected in the presence of heparin and intended for haematological and biochemical evaluation, under standardized conditions. The haematological evaluation consisted of red blood cell count, leukocyte counts, platelet counts, haematocrit, haemoglobin concentration, mean corpuscular volume (MCV), and mean corpuscular haemoglobin concentration (MCHC). The quantified biochemical parameters were: urea, creatinine, alanine aminotransaminase (ALT), aspartate aminotransaminase (AST) and alkaline phosphatase (ALP). While analysing the obtained data, it was possible to observe that only females presented divergences (P < 0.05) in the red blood cell series, in haemoglobin and in mean haemoglobin concentration (MCH). Regarding the analysis of the white blood cell series, females only presented differences (P < 0.05) in the leukocyte count. For males, there were variations (P < 0.05) in the counts of leukocytes, eosinophils, lymphocytes and neutrophils.[...]

Valores hematológicos e bioquímicos de camundongos imunossuprimidos BALB/c NUDE E C57BL/6 SCID / Haematological and biochemical values of immunosuppressed BALB/c NUDE and C57BL/6 SCID mice

Background: The emergence of the NUDE and SCID immunosuppressed mice lineages generated knowledge on various mechanisms of lymphocyte maturation and human autoimmune diseases. Information on haematological and biochemical parameters of these lineages is still scarce, making it impossible to infer homeostasis by comparing data, or to detect genetic influences on the parameters for these species. Haematological and biochemical tests were carried out on Balb/c NUDE and C57BL/6 SCID mice of both sexes, aiming to analyse the presence of genetic influence on possible variations of such parameters and to verify reference values for both lineages.Materials, Methods & Results: One hundred and forty mice (Mus musculus) of the Balb/C NUDE and C57BL/6 SCID lineages were used in the present study. The animals were previously anesthetized, the blood collection procedure was performed by cardiac puncture and the samples were collected in the presence of heparin and intended for haematological and biochemical evaluation, under standardized conditions. The haematological evaluation consisted of red blood cell count, leukocyte counts, platelet counts, haematocrit, haemoglobin concentration, mean corpuscular volume (MCV), and mean corpuscular haemoglobin concentration (MCHC). The quantified biochemical parameters were: urea, creatinine, alanine aminotransaminase (ALT), aspartate aminotransaminase (AST) and alkaline phosphatase (ALP). While analysing the obtained data, it was possible to observe that only females presented divergences (P < 0.05) in the red blood cell series, in haemoglobin and in mean haemoglobin concentration (MCH). Regarding the analysis of the white blood cell series, females only presented differences (P < 0.05) in the leukocyte count. For males, there were variations (P < 0.05) in the counts of leukocytes, eosinophils, lymphocytes and neutrophils.[...](AU)

Espermatogênese xenogênica pós-transplante de células tronco caninas no testículo de camundongos

As células tronco espermatogoniais (SSCs) são caracterizadas pela capacidade de autorrenovação, proliferação e transmissão das informações genéticas. Em caninos a primeira tentativa de xenotransplante não obteve o sucesso da produção de espermatozoides, no entanto, há evidências de que as células testiculares xenogênicas podem ser transplantadas no testículo do animal hospedeiro, e gerar espermatozoides viáveis do doador. Portanto, este estudo tem como objetivo realizar o xenotransplante das células germinativas caninas em camundongos imunosuprimidos, e com isto promover à produção de espermatozoides caninos viáveis, geneticamente modificados. E por meio desta técnica, analisar a eficiência da espermatogênese pós-transplante. Células germinativas testiculares foram caracterizadas, isoladas e cultivadas de cães pré-púberes, por meio de sistemas de cultura de enriquecimento e fatores de crescimento. As células foram transduzidas com um gene repórter GFP e LacZ, e por um vetor lentiviral para indentificar as SSCs nos testículos receptores. As SSCs transduzidas foram transplantadas nos testículos de camundongos (C57BL/6) tratados com Busulfan, após diferentes períodos os animais receptores foram eutanasiados e analisados. Aos 10 dias de cultivo as células germinativas adultas foram positivas para CD49f, CD117, e com 5 dias uma expressão semelhante de GFRA1 e DAZL, demonstrando a presença de SSCs e algumas células em meiose. Transplantamos 105 células e 20-43% das células transplantadas foram identificadas na membrana basal dos túbulos seminíferos do animal receptor. Portanto, o transplante das células germinativas caninas, mostrou que a purificação e o cultivo realizados são possíveis para obter SSCs caninas, as quais colonizaram os túbulos seminíferos dos camundongos imunodeficientes e mantiveram-se vivas na membrana basal por 90 dias após transplante, mesmo que estes animais tenham distância filogenética.

The spermatogonial stem cells (SSCs) are characterized by the capacity for self-renewal, proliferation and transmission of genetic information. In canines the first attempt to xenotransplantation did not achieve the success of sperm production. However, there is evidence that testicular xenogeneic cells can be transplanted into the testis of the host animal, and generate viable sperm donor. Therefore, this study aims conduct xenotransplantation of the canine germ cells in immunosuppressed mice, and thereby promote the production of viable sperm canines, genetically modified. And by this technique, analyze the efficiency of post-transplant spermatogenesis. Testicular germ cells were identified, isolated and cultured prepuberes dogs through enrichment culture systems and growth factors. Cells were modificated with a reporter gene GFP and LacZ. The SSCs canine was transplanted in mice (C57Bl/6), after different period and then the recipient animals were euthanized and analyzed. After 10 days in culture the germ cells were positive for CD49f, CD117, and 5 days a similar expression of GFRA1 and DAZL was observed, demonstrating the presence of SSCs and some cells in meiosis. 105 cells were transplanted and 20-43% of the transplanted cells were identified in the basement membrane of the seminiferous tubules after 90 days after transplantation. Therefore, the transplantation of canine germ cells showed that the cultivation and purification are performed possible for canine SSCs, it can colonize the seminiferous tubules the mice infertility remained alive in the basement membrane for 90 days after transplanting, even though these animals have phylogenetic distance.