MSCs derived from the umbilical cord tissue, termed UCX, were investigated for their immunomodulatory properties and compared to bone marrow-derived MSCs (BM-MSCs), the gold-standard in immunotherapy. Immunogenicity and immunosuppression were assessed by mixed lymphocyte reactions, suppression of lymphocyte proliferation and induction of regulatory T cells. Results showed that UCX were less immunogenic and showed higher immunosuppression activity than BM-MSCs. Further, UCX did not need prior activation or priming to exert their immunomodulatory effects. This was further corroborated in vivo in a model of acute inflammation. To elucidate the potency differences observed between UCX and BM-MSCs, gene expression related to immune modulation was analysed in both cell types. Several gene expression profile differences were found between UCX and BM-MSCs, namely decreased expression of HLA-DRA, HO-1, IGFBP1, 4 and 6, ILR1, IL6R and PTGES and increased expression of CD200, CD273, CD274, IL1B, IL-8, LIF and TGFB2. The latter were confirmed at the protein expression level. Overall, these results show that UCX seem to be naturally more potent immunosuppressors and less immunogenic than BM-MSCs. We propose that these differences may be due to increased levels of immunomodulatory surface proteins such as CD200, CD273, CD274 and cytokines such as IL1β, IL-8, LIF and TGFβ2.

IntroductionStandardization of mesenchymal stromal cells (MSCs) manufacturing is urgently needed to enable translational activities and ultimately facilitate comparison of clinical trial results. In this work we describe the adaptation of a proprietary method for isolation of a specific umbilical cord tissue-derived population of MSCs, herein designated by its registered trademark as UCX®, towards the production of an advanced therapy medicinal product (ATMP).MethodsThe adaptation focused on different stages of production, from cell isolation steps to cell culturing and cryopreservation. The origin and quality of materials and reagents were considered and steps for avoiding microbiological and endotoxin contamination of the final cell product were implemented. Cell isolation efficiency, MSCs surface markers and genetic profiles, originating from the use of different medium supplements, were compared. The ATMP-compliant UCX® product was also cryopreserved avoiding the use of dimethyl sulfoxide, an added benefit for the use of these cells as an ATMP. Cells were analyzed for expansion capacity and longevity. The final cell product was further characterized by flow cytometry, differentiation potential, and tested for contaminants at various passages. Finally, genetic stability and immune properties were also analyzed.ResultsThe isolation efficiency of UCX® was not affected by the introduction of clinical grade enzymes. Furthermore, isolation efficiencies and phenotype analyses revealed advantages in the use of human serum in cell culture as opposed to human platelet lysate. Initial decontamination of the tissue followed by the use of mycoplasma- and endotoxin-free materials and reagents in cell isolation and subsequent culture, enabled the removal of antibiotics during cell expansion. UCX®-ATMP maintained a significant expansion potential of 2.5 population doublings per week up to passage 15 (P15). They were also efficiently cryopreserved in a DMSO-free cryoprotectant medium with approximately 100% recovery and 98% viability post-thaw. Additionally, UCX®-ATMP were genetically stable upon expansion (up to P15) and maintained their immunomodulatory properties.ConclusionsWe have successfully adapted a method to consistently isolate, expand and cryopreserve a well-characterized population of human umbilical cord tissue-derived MSCs (UCX®), in order to obtain a cell product that is compliant with cell therapy. Here, we present quality and safety data that support the use of the UCX® as an ATMP, according to existing international guidelines.

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Cardiovascular disease is the leading cause of death in the set of chronic kidney disease (CKD) patients, whether on renal replacement therapy or conservative treatment. A better understanding of cardiovascular risk factors, diagnostic approach and management are central keys to develop strategies to reduce cardiovascular mortality among those patients. This review article discusses some aspects of pathophysiology, investigation methods and current treatment of cardiovascular disease in CKD patients.

-The usual drug used in the Wada test is amobarbital, but it is not available in Brazil. Propofol was already used by Bazin et al. in 1998, and in their report the test resulted good in the absence of any adverse effect. We report the use of propofol as the anesthetic for the Wada test. The test was carried out in a 26 years old woman with temporal medial lobe epilepsy refractory to medical treatment. Language functions and memory were tested after injection in both hemispheres by three procedures (Seattle, Montreal and Interview procedures). Propofol showed to be good to carry on the Wada test.KEY WORDS: Wada test, epilepsy, propofol, temporal lobe epilepsy. Teste de Wada com propofol em uma paciente com epilepsiaRESUMO -O amobarbital é a droga usada no teste de Wada, mas não é disponível em nosso pais. O propofol, usado por Bazin et al. em 1998, foi útil e sem efeitos adversos. Relatamos o uso do propofol como anestésico no teste de Wada. Este foi realizado como parte da avaliação pre-cirúrgica, em uma mulher de 26 anos com epilepsia do lobo temporal mesial, em uso de carbamazepina e ácido valpróico sem controle de suas crises. As funções da linguagem e memória foram testadas após injeção em ambos hemisférios separadamente por três procedimentos (Seattle, Montreal e Entrevista). O propofol mostrou-se eficaz para a realização do teste de Wada. PALAVRAS-CHAVE: teste de Wada, epilepsia, propofol, epilepsia do lobo temporal.