Protocol for isolating and assessing immunomodulatory ability of explant-cultured human amniotic membrane mesenchymal stem cells
Year: 2025
Type: article
Abstract: Mesenchymal stem cells (MSCs) are promising for regenerative medicine and cell-based therapies due to their multipotent and immunomodulatory properties. Here, we present a protocol for isolating MSCs from the amnion of the human amniotic membrane (AM-MSCs) using the explant culture approach and their subsequent characterization. We also describe steps for evaluating the immunomodulatory potential of the conditioned medium (CM) harvested from AM-MSCs, enabling assessment of its secretome without direct cell-to-cell contact. • Explant culture-based, non-enzymatic isolation of MSCs from human amniotic membrane • Reduced FBS concentration used for culture, compared to most protocols • Isolated AM-MSCs characterized by immunophenotyping and in vitro differentiation • Cell-free, CM-based assessment of the immunosuppressive potential of AM-MSCs Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Mesenchymal stem cells (MSCs) are promising for regenerative medicine and cell-based therapies due to their multipotent and immunomodulatory properties. Here, we present a protocol for isolating MSCs from the amnion of the human amniotic membrane (AM-MSCs) using the explant culture approach and their subsequent characterization. We also describe steps for evaluating the immunomodulatory potential of the conditioned medium (CM) harvested from AM-MSCs, enabling assessment of its secretome without direct cell-to-cell contact.
Source: STAR Protocols
Authors: Srishti Dutta Gupta, Pritam Banerjee, Tania Haoladar, Amar Aparajeet, Malancha Ta
Language: English
Subfield: Genetics
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $1,550
Award: G4956534364
OpenAlex ID: W4417535685