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Knee Osteoarthritis (OA knee) is a common degenerative disease particularly affecting the elderly population, resulting in limited mobility and impaired overall quality of life. Studies have shown that the pathogenesis of OA involves various factors, including inflammation, metabolic disorders, and mechanical forces. Moreover, synovial fluid in OA contains proinflammatory factors, leading to collagen damage and cartilage degradation. Adipose-derived stem cells (ADSCs) exhibit not only low immunogenicity characteristics but also participate in tissue repair and the restoration of biological functions by secreting secretomes, which result in the promotion of angiogenesis, immune modulation, and cell proliferation. This study aims to assess the therapeutic effects and potency of the allogeneic ADSC product ELIXCYTE® through an OA knee animal model. The OA knee animal model conducted on rats and rabbits, evaluated whether ELIXCYTE® improves knee activity. The potency analysis of ELIXCYTE® not only elucidates the anti-inflammatory mechanism but also identifies compelling biomarkers associated with efficacy, presenting new avenues for potential OA treatment. This study established OA animal models using both methods. In rat animal model, OA knee was induced using the Monosodium Iodoacetate (MIA), followed by an analysis of rat pain score, joint structure, and biomarkers. The injection of ELIXCYTE® led to a significant decrease in the knee pain, OARSI score, and an increase in type II collagen expression, along with decreased CD68 and MMP13 expression. The hematological analysis detected a reduction in the OA biomarker CTXII following ELIXCYTE® injection, suggesting that ELIXCYTE® can alleviate local inflammatory responses and protect cartilage integrity. Additionally, the anterior cruciate ligament transection (ACLT) animal model confirmed that the injection of ELIXCYTE® effectively increased type II collagen and reduced proinflammatory factors IL1β and TNFα. Significant improvements were also observed in MRI, histological, and ICRS scoring. Moreover, ELIXCYTE® expressed indoleamine 2,3 dioxygenase (IDO) in an inflammatory environment to inhibit T cell proliferation, with a positive correlation between IDO expression and T cell suppression. Also, coculturing ELIXCYTE® with naive macrophages reduced M1 polarization. In summary, ELIXCYTE® can improve the joint activity by modulating inflammatory responses and alleviating pain. [ABSTRACT FROM AUTHOR]