Work

Protocol for tendon stem/progenitor cell isolation from human tendon tissue

Year: 2025

Type: article

Abstract: Tendon stem/progenitor cells (TSPCs) are critical for tendon maintenance and repair, yet standardized approaches for their isolation and characterization remain limited. We present a protocol to isolate TSPCs from human tendons and establish them for in vitro culture. We describe steps for confirming phenotype and potency using a comprehensive marker panel through flow cytometry, qPCR, immunofluorescence, and colorimetric staining. This approach consistently yields a homogeneous TSPC population, enabling reproducible in vitro studies of tendon biology and applications in regenerative therapies. • Steps for isolating tendon stem/progenitor cells from human tendon tissue • Procedure for freezing, reviving, and expanding isolated cells in culture • Confirm phenotype using flow cytometry, immunofluorescence, and RT-qPCR • Confirm potency by the cells’ ability to form adipocytes and osteoblasts Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Tendon stem/progenitor cells (TSPCs) are critical for tendon maintenance and repair, yet standardized approaches for their isolation and characterization remain limited. We present a protocol to isolate TSPCs from human tendons and establish them for in vitro culture. We describe steps for confirming phenotype and potency using a comprehensive marker panel through flow cytometry, qPCR, immunofluorescence, and colorimetric staining. This approach consistently yields a homogeneous TSPC population, enabling reproducible in vitro studies of tendon biology and applications in regenerative therapies.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.104143