Efficient in vitro expansion of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) requires biocompatible, xenofree culture surfaces. Here, Tamarindus indica L. seed kernel powder (TKP) was evaluated as a natural, cost-effective alternative to xenogenic gelatin. Preliminary phytochemical screening and GC–MS profiling confirmed bioactive compounds in TKP. FTIR and XRD analyses revealed functional groups and structural differences compared to gelatin (GEL) and regular tissue culture dishes (TTP), supporting enhanced hUC-MSC adhesion and proliferation. SEM demonstrated the favourable surface roughness of TKP coatings. DPPH assay showed 51.35% radical scavenging at 1 mg/mL with an IC₅₀ of 0.96 mg/mL. MTT assay indicated 90% cell viability at 2% TKP. Optimized TKP coating (2%, 0.1 mL/cm² area) enabled efficient expansion at passages P0–P2, yielding 0.7 × 10⁶ cells/mL (95% viability) on TKP and 0.82 × 10⁶ cells/mL (94% viability) on GEL. AO/EtBr staining confirmed higher viable cell numbers on TKP against GEL and TTP. Flow cytometry (CD90⁺, CD105⁺, CD31⁻) and RT-PCR (CD90, CD73, CD105) verified retention of hUC-MSC markers. All experiments were performed in triplicate; results are mean ± SD, with p < 0.05. Culturing hUC-MSCs on TKP in human platelet lysate (HPL) media establishes a fully xenofree system. Collectively, TKP represents a sustainable, bioactive coating for scalable hUC-MSC culture, offering a promising alternative to conventional gelatin-based surfaces for regenerative applications. Although TKP enables efficient hUC-MSC expansion, potential batch-to-batch variability, unexamined protein expression under diverse culture conditions, and the absence of long-term or in vivo validation highlight the need for further studies.
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