To assess current pediatric cholesterol screening practices, and attitudes, among pediatric primary care providers (PCPs) via qualitative semistructured interviews designed to identify barriers and facilitators to universal cholesterol screening practices recommended by the National Heart Blood and Lung Institute and the American Academy of Pediatrics. An online survey and subsequent 30-minute semistructured phone interview were completed with PCPs from regions in Northern California and Minnesota (survey n = 25, interview n = 12). Interviews were qualitatively analyzed using the consolidated framework for implementation research to categorize barriers, facilitators, and strategies to increase pediatric cholesterol screening among PCPs. PCPs from California (n = 8) and Minnesota (n = 4) consistently identified cost of cholesterol screening, particularly the cost of time due to competing visit priorities, as a barrier. A supportive learning environment, feelings of self-efficacy, access to resources, and well-established clinical networks with specialists (eg, cardiologists) were facilitators to screening. The perceived level of endorsement behind cholesterol screening within the clinic, perceived validity of national guidelines, and ability to adapt guidelines to existing clinical workflow were notable differentiators between high vs low self-reported screen rates. Findings of this study suggest that efforts to increase universal pediatric cholesterol screening will likely require the development of strategies to increase provider education about the long-term benefits of cholesterol screening (knowledge and beliefs), and ensuring providers feel supported and empowered when assessing/acting on the results of this screening (self-efficacy, engaging leaders, networks, and communication).