The culture and identification of human placenta-derived mesenchymal stem cells

Objective: This study aimed to establish a reliable protocol for the cultivation and characterization of human placenta-derived mesenchymal stem cells (hPMSCs) to evaluate their growth dynamics and immunophenotype. Methods: hPMSCs were thawed and cultured under controlled conditions using specialized serum-free medium. Cell viability and morphology were assessed using an inverted microscope, and medium changes were performed bi-daily. For cell identification, immunofluorescence staining was conducted with antibodies CD44, CD90, and CD45, and cells were characterized based on surface marker expression. Results: Cultured hPMSCs exhibited a fibroblast-like morphology with rapid proliferation, particularly after reaching a seeding density of 50%. Growth curves indicated peak proliferation between days 3 and 4. Immunofluorescence analysis confirmed that hPMSCs were positive for CD90 and CD44, but negative for CD45, aligning with typical mesenchymal stem cell profiles. Conclusion: The established protocol successfully cultivated and characterized hPMSCs, demonstrating their viability and specific surface markers. These findings support their potential application in regenerative medicine and therapeutic research.