RESUMO
BACKGROUND AIMS: Human mesenchymal stromal cells (hMSCs) and their secreted products show great promise for treatment of musculoskeletal injury and inflammatory or immune diseases. However, the path to clinical utilization is hampered by donor-tissue variation and the inability to manufacture clinically relevant yields of cells or their products in a cost-effective manner. Previously we described a method to produce chemically and mechanically customizable gelatin methacryloyl (GelMA) microcarriers for culture of hMSCs. Herein, we demonstrate scalable GelMA microcarrier-mediated expansion of induced pluripotent stem cell (iPSC)-derived hMSCs (ihMSCs) in 500 mL and 3L vertical wheel bioreactors, offering several advantages over conventional microcarrier and monolayer-based expansion strategies. METHODS: Human mesenchymal stromal cells derived from induced pluripotent cells were cultured on custom-made spherical gelatin methacryloyl microcarriers in single-use vertical wheel bioreactors (PBS Biotech). Cell-laden microcarriers were visualized using confocal microscopy and elastic light scattering methodologies. Cells were assayed for viability and differentiation potential in vitro by standard methods. Osteogenic cell matrix derived from cells was tested in vitro for osteogenic healing using a rodent calvarial defect assay. Immune modulation was assayed with an in vivo peritonitis model using Zymozan A. RESULTS: The optical properties of GelMA microcarriers permit noninvasive visualization of cells with elastic light scattering modalities, and harvest of product is streamlined by microcarrier digestion. At volumes above 500 mL, the process is significantly more cost-effective than monolayer culture. Osteogenic cell matrix derived from ihMSCs expanded on GelMA microcarriers exhibited enhanced in vivo bone regenerative capacity when compared to bone morphogenic protein 2, and the ihMSCs exhibited superior immunosuppressive properties in vivo when compared to monolayer-generated ihMSCs. CONCLUSIONS: These results indicate that the cell expansion strategy described here represents a superior approach for efficient generation, monitoring and harvest of therapeutic MSCs and their products.
Assuntos
Técnicas de Cultura de Células , Células-Tronco Mesenquimais , Humanos , Técnicas de Cultura de Células/métodos , Reatores Biológicos , Osteogênese , Regeneração Óssea , Proliferação de Células , Diferenciação Celular , Células CultivadasRESUMO
Food insecurity on college campuses is a major public health problem and has been documented for the last decade. Sufficient food access is a crucial social determinant of health, thus campuses across the country have implemented various programmes, systems and policies to enhance access to food which have included food pantries, campus gardens, farmers' markets, meal share or voucher programmes, mobile food applications, campus food gleaning, food recovery efforts, meal deliveries and task force/working groups. However, little is understood about how to best address food insecurity and support students who are struggling with basic needs. The impact of food insecurity on students' academic and social success, in addition to their overall well-being, should be investigated and prioritised at each higher education institution. This is especially true for marginalised students, such as minority or first-generation students, who are at heightened risk for food insecurity. In order to create a culture of health equity, in which most at-risk students are provided resources and opportunities to achieve optimal well-being, higher education institutions must prioritise mitigating food insecurity on the college campus. Higher education institutions could benefit from adopting comprehensive and individualised approaches to promoting food security for marginalised students in order to facilitate equal opportunity for optimal scholastic achievement among students of all socio-demographic backgrounds.
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Human mesenchymal stem cells (hMSCs) are effective in treating disorders resulting from an inflammatory or heightened immune response. The hMSCs derived from induced pluripotent stem cells (ihMSCs) share the characteristics of tissue derived hMSCs but lack challenges associated with limited tissue sources and donor variation. To meet the expected future demand for ihMSCs, there is a need to develop scalable methods for their production at clinical yields while retaining immunomodulatory efficacy. Herein, we describe a platform for the scalable expansion and rapid harvest of ihMSCs with robust immunomodulatory activity using degradable gelatin methacryloyl (GelMA) microcarriers. GelMA microcarriers were rapidly and reproducibly fabricated using a custom microfluidic step emulsification device at relatively low cost. Using vertical wheel bioreactors, 8.8 to 16.3-fold expansion of ihMSCs was achieved over 8 days. Complete recovery by 5-minute digestion of the microcarriers with standard cell dissociation reagents resulted in >95% viability. The ihMSCs matched or exceeded immunomodulatory potential in vitro when compared with ihMSCs expanded on monolayers. This is the first description of a robust, scalable, and cost-effective method for generation of immunomodulatory ihMSCs, representing a significant contribution to their translational potential.
Assuntos
Células-Tronco Pluripotentes Induzidas , Células-Tronco Mesenquimais , Reatores Biológicos , Técnicas de Cultura de Células/métodos , Diferenciação Celular , Proliferação de Células , Gelatina/farmacologia , Humanos , MetacrilatosRESUMO
The purpose of this study was to examine the impact of a sport nutrition education and life-skills intervention on sport nutrition knowledge (SNK), attitudes/beliefs and dietary behaviors relevant to sport nutrition among high school (HS) soccer players. Three assessments were done over the 2-year intervention (baseline = time 1, end year 1 = time 2, end year 2 = time 3). Participants (n = 217; females = 64%; Latino = 47.5%; 14.9 ± 0.9-year; 46.5% National School Breakfast/Lunch Program) were assigned to an intervention group (IG, n = 153; 9 schools) or comparison group (CG, n = 64; 4 schools) based on geographical location. Differences over time were examined based on group, sex, socioeconomic status (SES) and race/ethnicity. The IG increased SNK scores by ~10% (time 1 = 51.6%; time 3 = 60.9%; p ≤ 0.001), with the greatest change in the female IG vs. CG and no differences in male IG vs. CG. Daily breakfast consumption was 53.7% in both groups. IG players were 3 times more likely (95%CI = 2.59, 7.77) to report trying to eat for performance (IG = 48.7% vs. CG = 30.2%). By time 3, IG players were less likely to report that 'diet met nutritional requirements' (31.6%) compared to CG (47.6%). For IG, the consumption of lunch (≥5-days/week) did not change (92.2â»93.4%), but declined in the CG (90.6%) (p = 0.04). No other differences by sub-population (race/ethnicity, SES) were observed. Our findings indicate that HS athletes are motivated to learn and improve diet behaviors, and benefit from team-based nutrition interventions. Future interventions should consider delivery of curriculum/experiential learning during a defined training period, with messages reinforced with supports at home, school and athletic settings.
Assuntos
Fenômenos Fisiológicos da Nutrição do Adolescente , Conhecimentos, Atitudes e Prática em Saúde , Futebol , Ciências da Nutrição e do Esporte , Adolescente , Feminino , Humanos , Masculino , Estudantes , Adulto JovemRESUMO
The purpose of this study was to compare changes in diet and daily physical activity (PA) in high school (HS) soccer players who participated in either a two-year obesity prevention intervention or comparison group, while controlling for sex, race/ethnicity, and socioeconomic status. Participants (n = 388; females = 58%; Latino = 38%; 15.3 ± 1.1 years, 38% National School Breakfast/Lunch Program) were assigned to either an intervention (n = 278; 9 schools) or comparison group (n = 110; 4 schools) based on geographical location. Pre/post intervention assessment of diet was done using Block Fat/Sugar/Fruit/Vegetable Screener, and daily steps was done using the Fitbit-Zip. Groups were compared over-time for mean changes (post-pre) in fruit/vegetables (FV), saturated fat (SF), added sugar, and PA (daily steps, moderate-to-vigorous PA) using analysis of covariance. The two-year intervention decreased mean added sugar intake (-12.1 g/day, CI (7.4, 16.8), p = 0.02); there were no differences in groups for FV or SF intake (p = 0.89). For both groups, PA was significantly higher in-soccer (9937 steps/day) vs. out-of-soccer season (8117 steps/day), emphasizing the contribution of organized sports to youth daily PA. At baseline, Latino youth had significantly higher added sugar intake (+14 g/day, p < 0.01) than non-Latinos. Targeting active youth in a diet/PA intervention improves diet, but out of soccer season youth need engagement to maintain PA (200).
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Atletas , Dieta Saudável , Exercício Físico , Obesidade Infantil , Instituições Acadêmicas , Adolescente , Comportamento do Adolescente , Feminino , Comportamentos Relacionados com a Saúde , Conhecimentos, Atitudes e Prática em Saúde , Promoção da Saúde , Humanos , Masculino , Avaliação de Programas e Projetos de Saúde , Serviços de Saúde EscolarRESUMO
This paper reports the process data on program fidelity, best practices for intervention implementation, youth and coach engagement, and youth application of knowledge and skills for the two-year WAVE~Ripples for Change (WAVE) obesity prevention intervention program focused on healthy eating, physical activity, and life skills with high school (HS) soccer players aged 14â»19 years. Internal (staff: n = 7; volunteers: n = 27) and external (youth: n = 100; coaches: n = 9) stakeholders were interviewed/ surveyed. Staff rated program fidelity as high (94%), as did volunteers (85%). Best practices included coach encouragement for athlete participation, use of on-line consent for enrollment, building relationships with HS staff to complete assessments, sending text reminders, and providing incentives. Study results showed an enrollment rate of 72%, completion of baseline assessments of 89â»98%, attendance of sports nutrition lessons in Year 1 and Year 2 of 90% and 39%, respectively, and team-building workshop (TBW) attendance of 25â»31%. Activities exceeding youth expectations (>90%) included, (1) activities with their soccer team; (2) the TBW-cooking; and (3) sports nutrition lessons. The obesity prevention skills most applied by youth were obtained from the TBW-gardening and harvesting (49%), the TBW-cooking (43%), and sports nutrition lessons (44%). Coaches also rated the sports nutrition lessons highly and reported increased awareness for hydration/fueling during sport by the athletes. Using sport teams/clubs to engage youth in obesity prevention is a feasible model for future study.