Resumo
Stem cell therapy has shown promising results in tendinitis and osteoarthritis in equine medicine. The purpose of this work was to characterize the adipose-derived mesenchymal stem cells (AdMSCs) in horses through (1) the assessment of the capacity of progenitor cells to perform adipogenic, osteogenic and chondrogenic differentiation; and (2) flow cytometry analysis using the stemness related markers: CD44, CD90, CD105 and MHC Class II. Five mixed-breed horses, aged 2-4 years-old were used to collect adipose tissue from the base of the tail. After isolation and culture of AdMSCs, immunophenotypic characterization was performed through flow cytometry. There was a high expression of CD44, CD90 and CD105, and no expression of MHC Class II markers. The tri-lineage differentiation was confirmed by specific staining: adipogenic (Oil Red O), osteogenic (Alizarin Red), and chondrogenic (Alcian Blue). The equine AdMSCs are a promising type of adult progenitor cell for tissue engineering in veterinary medicine.(AU)
O uso de células tronco tem demonstrado resultados promissores na terapia da tendinite e osteoartrite na medicina equina. O objetivo deste trabalho foi caracterizar as células tronco mesenquimais derivadas do tecido adiposo (AdCTMs) em cavalos através da (1) avaliação da capacidade das células progenitoras para realizar a diferenciação adipogênica, osteogênica e condrogênica; e (2) através da análise por citometria de fluxo, utilizando os marcadores stemness relacionados: CD44, CD90, CD105 e MHC de Classe II. Cinco cavalos sem raça definida, de 2 a 4 anos de idade foram utilizados para a coleta do tecido adiposo da base da cauda. Após o isolamento e cultivo das AdCTMs, a caracterização imunofenotípica foi realizada pela citometria de fluxo. Houve alta expressão dos marcadores CD44, CD90 e CD105, e não houve expressão do MHC Classe II. A diferenciação foi confirmada pela coloração específica: adipogênica (Oil Red O), osteogênico (Alizarin Red), e condrogênico (Alcian Blue). As AdCTMs são um tipo promissor de células progenitoras adulta para a engenharia tecidual na medicina veterinária.(AU)
Assuntos
Animais , Células-Tronco/citologia , Tecido Adiposo/fisiologia , Imunofenotipagem , Cavalos/classificaçãoResumo
A cross-sectional study was performed on HIV-1 infected individuals with or without antiretroviral treatment (ARV) in the AIDS Day Hospital, Botucatu Medical School, UNESP. Between August 2004 and October 2005, 73 HIV-1 infected individuals were divided into three groups: infected individuals with or without AIDS who had never received ARV (G1 = 15); patients on HAART that had had plasma HIV-1 RNA viral load (VL) equal to or greater than 50 copies/mL (G2 = 27); and patients on HAART with undetectable VL for at least the past six months (G3 = 31). There was also an additional group that comprised blood donors without any sign of the disease and with negative HIV serum tests (G4 = 20), which was the control group. Serum cytokine levels (values in pg/mL) were measured by enzyme-linked immunosorbent assay (ELISA) and specific mRNA expression by reverse transcription polymerase chain reaction (RT-PCR). Both techniques were performed on the four groups for TNF-á, IL-2, INF-ã, IL-4 and IL-10. All patients were submitted to VL determination and CD4+ and CD8+T lymphocyte counts. The analysis of the results revealed a significant comparison among groups for both methods and an association between the latter (> 80% r² > 0.80). There was only one exception, in control individuals for IL-2 by ELISA. The cytokine profiles, in both methods, for the three patient groups, were mature Th-0. The behaviors of IL-2 and INF-ã required emphasis due to consequent expression of dominant Th profile. Both methods showed low IL-2 and high mean values of INF-ã in the three groups. Several authors have recently drawn attention to the substantial apoptosis of infected and non-infected CD4+T cells, mainly during primary infection, persisting only in those with INF-ã phenotype producer and not IL-2. HIV infected individuals submitted to HAART are expected to produce IL-2 in an attempt to present Th-1 profile, but in most cases this did not occur.(AU)