Adipose tissue-derived stem cell therapy in rat cryopreserved ovarian grafts

Abstract\ud The preliminary results of ovarian transplantation in clinical practice are encouraging. However, the follicular depletion caused by ischemic injury is a main concern and is directly related to short-term graft survival. Cell therapy with adipose tissue-derived stem cells (ASCs) could be an alternative to induce early angiogenesis in the graft. This study aimed to evaluate ASCs therapy in rat cryopreserved ovarian grafts. A single dose of rat ASC (rASCs) or vehicle was injected into the bilateral cryopreserved ovaries of twelve adult female rats immediately after an autologous transplant. Daily vaginal smears were performed for estrous cycle evaluation until euthanasia on postoperative day 30. Follicle viability, graft morphology and apoptosis were assessed. No differences were found with respect to estrous cycle resumption and follicle viability (P > 0.05). However, compared with the vehicle-treated grafts, the morphology of the ASCs-treated grafts was impaired, with diffuse atrophy and increased apoptosis (P < 0.05). ASCs direct injected in the stroma of rat cryopreserved ovarian grafts impaired its morphology although may not interfere with the functional resumption on short-term. Further investigations are necessary to evaluated whether it could compromise their viability in the long-term.We thank: 1) São Paulo Research Foundation (FAPESP) for grant support\ud (Process numbers: 2010/17897-5 and 2012/09469-9) and Coordenação de\ud Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for post-doctoral\ud scholarship

Cell therapy for ischemic cardiac disease: effect of different routes for cell administration, time post-mi and the use of a fibrin polymer for cardiac cell retention and myocardial function

A terapia celular representa uma abordagem promissora para o tratamento de cardiopatia isquêmica, porém aspectos-chave dessa estratégia permanecem incertos. Neste trabalho avaliamos a eficiência da retenção cardíaca de células da medula óssea marcadas com tecnécio (99m Tc-CMO) transplantadas, de acordo com o tempo após o infarto (1, 2, 3 e 7 dias) e a via de administração dessas células (intravenosa [IV], intraventricular [IC], intracoronariana [ICO] e intramiocárdica [IM]), em ratos submetidos à isquemia-reperfusão cardíaca [I&R]. Após 24 horas, a retenção cardíaca de 99m Tc-CMO foi maior na via IM comparada com a média alcançada pelas demais (6,79% do total injetado vs. 0,53%). O uso de fibrina como veículo para a injeção de células incrementou a retenção em 2.5 vezes (17,12 vs. 6,84%) na via IM. Curiosamente, quando administradas após 7 dias, a retenção de células na via IM alcançou valores próximos dos observados com da matriz de fibrina injetadas 24 h após a I&R (16,55 vs. 17,12%), enquanto que para as demais vias as mudanças foram insignificantes. Nos animais em que as CMO foram administradas por via intramiocárdica 24 horas após a I&R, com ou sem fibrina, observou-se melhora significante do desempenho cardíaco frente ao estresse farmacológico com fenilefrina quando comparados aos controles. Em conjunto, os dados mostram a biodistribuição das células injetadas após a I&R por 4 diferentes vias e 4 intervalos de tempo pós-lesão e indicam que a via IM é a que produz maior retenção cardíaca. O uso do biopolímero de fibrina aumenta a retenção das células e a eficácia deste efeito sobre a função cardíaca e mortalidade dos animais em longo prazo, além de 30 dias pós I&R, merecerá ser investigada no futuro.Cell therapy represents a promising approach for ischemic cardiac disease, but key aspects of this strategy remain unclear. We examined the effects of timing and route of administration of bone marrow cells (BMCs) after myocardial ischemia/reperfusion injury (I&R). 99mTc-labeled BMCs were injected by 4 different routes: intravenous (IV), left ventricular cavity (LV), left ventricular cavity with temporal aorta occlusion (LV+) and intramyocardial (IM). The injections were performed 1, 2, 3, or 7 days after infarction. Cardiac retention was higher following the IM route compared to the average values obtained by all other routes (6.79% of the total radioactivity injected vs. 0.53%). Use of a fibrin biopolymer as vehicle during IM injection led to a 2.5-fold increase in cardiac cell accumulation (17.12 vs. 6.84%). Interestingly, the retention of cells administered with culture medium at day 7 post-MI by the IM route was similar to that observed when cells were injected 24 h post-IM using fibrin (16.55 vs 17.12%), whereas no significant changes were observed for the other routes. Cell therapy 24 hs post MI by IM injection, with or without fibrin, resulted in comparable improvement in cardiac function under pharmacological stress compared to control animals. Together, we provide evidence for the biodistribution of 99mTc-labeled BMCs injected post MI by 4 different routes and times post-injury, which shows that the IM rout is the most effective for cardiac cell retention. The use of a fibrin biopolymer further increased cardiac cell retention and its potential long term benefits, beyond 30, on reducing mortality and improving cardiac function deserve to be explored in the future

Implications for Clinical Practice from a Multicenter Survey of Heart Failure Management Centers

OBJECTIVES: This observational, cross-sectional study based aimed to test whether heart failure (HF)-disease management program (DMP) components are influencing care and clinical decision-making in Brazil. METHODS: The survey respondents were cardiologists recommended by experts in the field and invited to participate in the survey via printed form or email. The survey consisted of 29 questions addressing site demographics, public versus private infrastructure, HF baseline data of patients, clinical management of HF, performance indicators, and perceptions about HF treatment. RESULTS: Data were obtained from 98 centers (58% public and 42% private practice) distributed across Brazil. Public HF-DMPs compared to private HF-DMP were associated with a higher percentage of HF-DMP-dedicated services (79% vs 24%; OR: 12, 95% CI: 94-34), multidisciplinary HF (MHF)-DMP [84% vs 65%; OR: 3; 95% CI: 1-8), HF educational programs (49% vs 18%; OR: 4; 95% CI: 1-2), written instructions before hospital discharge (83% vs 76%; OR: 1; 95% CI: 0-5), rehabilitation (69% vs 39%; OR: 3; 95% CI: 1-9), monitoring (44% vs 29%; OR: 2; 95% CI: 1-5), guideline-directed medical therapy-HF use (94% vs 85%; OR: 3; 95% CI: 0-15), and less B-type natriuretic peptide (BNP) dosage (73% vs 88%; OR: 3; 95% CI: 1-9), and key performance indicators (37% vs 60%; OR: 3; 95% CI: 1-7). In comparison to non- MHF-DMP, MHF-DMP was associated with more educational initiatives (42% vs 6%; OR: 12; 95% CI: 1-97), written instructions (83% vs 68%; OR: 2: 95% CI: 1-7), rehabilitation (69% vs 17%; OR: 11; 95% CI: 3-44), monitoring (47% vs 6%; OR: 14; 95% CI: 2-115), GDMT-HF (92% vs 83%; OR: 3; 95% CI: 0-15). In addition, there were less use of BNP as a biomarker (70% vs 84%; OR: 2; 95% CI: 1-8) and key performance indicators (35% vs 51%; OR: 2; 95% CI: 91,6) in the non-MHF group. Physicians considered changing or introducing new medications mostly when patients were hospitalized or when observing worsening disease and/or symptoms. Adherence to drug treatment and non-drug treatment factors were the greatest medical problems associated with HF treatment. CONCLUSION: HF-DMPs are highly heterogeneous. New strategies for HF care should consider the present study highlights and clinical decision-making processes to improve HF patient care

Porcine Adipose Tissue-Derived Mesenchymal Stem Cells Retain Their Proliferative Characteristics, Senescence, Karyotype and Plasticity after Long-Term Cryopreservation

<div><p>We and others have provided evidence that adipose tissue-derived mesenchymal stem cells (ASCs) can mitigate rat cardiac functional deterioration after myocardial ischemia, even though the mechanism of action or the relevance of these findings to human conditions remains elusive. In this regard, the porcine model is a key translational step, because it displays heart anatomic-physiological features that are similar to those found in the human heart. Towards this end, we wanted to establish the cultural characteristics of porcine ASCs (pASCs) with or without long-term cryostorage, considering that allogeneic transplantation may also be a future option. Compared to fresh pASCs, thawed cells displayed 90–95% viability and no changes in morphological characteristics or in the expression of surface markers (being pASCs characterized by positive markers CD29⁺; CD90⁺; CD44⁺; CD140b⁺; CD105⁺; and negative markers CD31⁻; CD34⁻; CD45⁻ and SLA-DR⁻; n = 3). Mean population doubling time was also comparable (64.26±15.11 hours to thawed cells vs. 62.74±18.07 hours to fresh cells) and cumulative population doubling increased constantly until Passage 10 (P10) in the entire cell population, with a small and gradual increase in senescence (P5, 3.25%±0.26 vs. 3.47%±0.32 and P10, 9.6%±0.29 vs. 10.67%±1.25, thawed vs. fresh; SA-β-Gal staining). Chromosomal aberrations were not observed. In addition, under both conditions pASCs responded to adipogenic and osteogenic chemical cues <i>in vitro</i>. In conclusion, we have demonstrated the growth characteristics, senescence, and the capacity of pASCs to respond to chemical cues <i>in vitro</i> and have provided evidence that these properties are not influenced by cryostorage in 10% DMSO solution.</p></div

Cryostorage does not influence the expression of mesenchymal surface markers.

<p><b>A)</b> FACS histogram representations for each analyzed surface marker in pASCs. <b>B)</b> Quantification of surface markers (means expressed as percentages; n = 3). <b>C)</b> Representative semi quantitative RT-PCR comparing fresh and thawed cells by mRNA expression for positive and negative phenotypic markers (2% agarose gel). Numbers 1, 2 and 3 are relative to different animals used to extract the pASCs.</p