Modified Isolation Method of Mesenchymal Stem Cells from Placental Tissue: Cheap and Effective

Mesenchymal stem cells (MSCs) originating from the human placenta (PMSCs) are promising for cell-based therapeutics. However, the growing demand for PMSCs in clinical trials necessitates high-quality cells in huge numbers. This study aims to create an effective and cheap procedure for PMSC isolation and culture. MSC was isolated from human placental tissue (3- 4mm) by three different methods: explant, collagenase (1 mg/ml), modified method (explant+ 0.1 mg/ml collagenase). PMSC cell morphologies were followed under an inverted microscope for 21 days. PMSCs characterizations were performed using CD44 and CD90 staining and the immunofluorescence method. PMSCs differentiation capacities were determined by alcian blue, oil red, and alizarin red staining. The modified method (explant+collagenase) is based on placenta tissue fragments put in the bottom of the dish and incubated with a culture medium containing %0,1 collagenase type 1. Compared to the traditional explant and enzymatic culture methods that used collagenase 1 mg/ml for incubation in terms of isolation efficiency, cell yield, and proliferative capability. Immunofluorescence staining demonstrated the characterization of mesenchymal stem cells for all isolation techniques. It was determined that the number of cells per area was the lowest in the explant method and the highest in the modified method. Morphological structures of PMSCs isolated using explant, collagenase, and modified method are fusiform and have a fibroblast-like appearance. PMSC isolated in the modified method has higher quality cells with high proliferation ability. It was observed that the modified method especially preserved the cell adhesion ability. It was observed that the isolated PMSCs using a modified method could differentiate into adipogenic, chondrogenic, and osteogenic cell lines in the stainings performed to evaluate their differentiation capacity. Our research identified an effective and high-yield method for producing high-quality PMSCs from the placenta tissue. According to general isolation protocols, the low amount of enzyme used makes it an ideal isolation technique for MSCs for therapeutic use.