Adipose-derived stem cells (ADSCs) are used in tissue regeneration therapies. The objective of this study is to identify stable reference genes (RGs) for use in gene expression studies in a characterized equine adipose-derived mesenchymal stem cell (EADMSC) differentiation model. ADSCs were differentiated into adipocytes (ADs) or osteoblasts (OBs), and the proteomes from these cells were analyzed by liquid chromatography tandem mass spectrometry. Proteins that were stably expressed in all three cells types were identified, and the mRNA expression stabilities for their corresponding genes were validated by RT-qPCR. PPP6R1, CCDC97, and then either ACTB or EPHA2 demonstrated the most stable mRNA levels. Normalizing target gene C_q data with at least three of these RGs simultaneously, as per MIQE guidelines (PPP6R1 and CCDC97 with either ACTB or EPHA2), resulted in congruent conclusions. FABP5 expression was increased in ADs (5.99 and 8.00 fold, p = 0.00002 and p = 0.0003) and in OBs (5.18 and 5.91 fold, p = 0.0011 and p = 0.0023) relative to ADSCs. RUNX2 expression was slightly higher in ADs relative to ADSCs (1.97 and 2.65 fold, p = 0.04 and p = 0.01), but not in OBs (0.9 and 1.03 fold, p = 0.58 and p = 0.91).

The aim of this research was to determine the impact of heat stress on cell differentiation in an equine mesenchymal stem cell model (EMSC) through the application of heat stress to primary EMSCs as they progressed through the cell specialization process. A proteomic analysis was performed using mass spectrometry to compare relative protein abundances among the proteomes of three cell types: progenitor EMSCs and differentiated osteoblasts and adipocytes, maintained at 37 °C and 42 °C during the process of cell differentiation. A cell-type and temperature-specific response to heat stress was observed, and many of the specific differentially expressed proteins were involved in cell-signaling pathways such as Notch and Wnt signaling, which are known to regulate cellular development. Furthermore, cytoskeletal proteins profilin, DSTN, SPECC1, and DAAM2 showed increased protein levels in osteoblasts differentiated at 42 °C as compared with 37 °C, and these cells, while they appeared to accumulate calcium, did not organize into a whorl agglomerate as is typically seen at physiological temperatures. This altered proteome composition observed suggests that heat stress could have long-term impacts on cellular development. We propose that this in vitro stem cell culture model of cell differentiation is useful for investigating molecular mechanisms that impact cell development in response to stressors.

BackgroundGovernments, private industries and not-for-profit organizations are increasingly investing resources in support programs for patients. These programs may provide self-management strategies enabling patients to manage their disease and medication regimen and/or access to medications, therapies and health services, with the goal to improve health outcomes. The objectives of this scoping review were to: (i) describe the characteristics of patient support programs (PSPs), for patients on medication regimens, (ii) identify subthemes that describe PSP services, and (iii) to classify the service subthemes, using an integrated people-centered (IP-C) perspective. MethodsPRISMA-ScR methodology and Covidence software were used to facilitate the review of 479 articles, and the selection and screening of 70 original research articles that described PSP services and their impact on clinical, humanistic and/or economic outcomes. A search of PubMed, Embase, CINAHL, Academic Search Ultimate, and Web of Science was conducted covering a 10 year period, from 2010 to 2020. The PSPs identified were organized into subthemes identified as part of an inductive process. The thematic areas were grouped into one of three people-centered categories that reflect the WHO vision for IPC health care. ResultsThe 70 articles reviewed, describe PSPs covering more than nine disease categories. The majority of the articles were published in North America, predominantly in the United States (26) and Canada (11), and also in Germany (6), the United Kingdom (6), China (4) and other countries (18). PSPs were funded and delivered by private drug or device manufacturers, government organizations, specialty pharmacies and also non-governmental groups (e.g., not-for-profit organizations), private hospitals, private payers (e.g., insurance companies) and various partnership types among these organizations. Fifty-six unique PSPs were identified that offered a range of services. Twenty-five PSP service subthemes were identified and these were each placed into one of three over-arching patient-centered categories: (i) access and equity services; (ii) quality and health services, and (iii) patient and caregiver empowerment and self-management services.ConclusionsPSPs offer various services that can be optimized and synergized to facilitate equitable, accessible, quality health services that support patient empowerment for the self-management of disease for improved health outcomes.