Comparison of Efficiency of Self-renewal and Differentiation Potential in Tendon-derived Mesenchymal Stem Cells Isolated by Magnetic-activated Cell Sorting Method or Colony Picking Method.
10.14193/jkfas.2014.18.3.100
- Author:
Moses LEE
1
;
Yoorim CHOI
;
Dong Suk YOON
;
Jin Woo LEE
;
Gil Sung YOON
;
Woo Jin CHOI
;
Seung Hwan HAN
Author Information
1. Department of Orthopaedic Surgery, Yonsei University College of Medicine, Seoul, Korea. OSMEDIC@yuhs.ac
- Publication Type:In Vitro ; Original Article
- Keywords:
Tendons;
Mesenchymal stem cells;
Magnetic-activated cell sorting;
Colony picking method
- MeSH:
Clone Cells;
Cloning, Organism;
Digestion;
Flow Cytometry;
Humans;
Mesenchymal Stromal Cells*;
Tendons;
Tissue Donors
- From:Journal of Korean Foot and Ankle Society
2014;18(3):100-107
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
PURPOSE: The purpose of this study is to evaluate the efficacy of mesenchymal stem cell (MSC) isolation by the magnetic-activated cell sorting (MACS) method in tendon tissue-derived cells compared to the colony picking method for isolation of MSCs by picking colonyforming cells. MATERIALS AND METHODS: Human tendon-derived cells were isolated by enzyme digestion using normal tendon tissues from three donors. We used the magnetic kit and well-known MSC markers (CD90 or CD105) to isolate MSCs in tendon-derived cells using MACS. Cloning cylinders were used to isolate colony-forming cells having MSC characteristics in tendon-derived cells. Colony-forming unitfibroblast (CFU-F) assay was used to evaluate the self-renewal capacity of cells isolated using the colony picking method or MACS. For comparison of differentiation potentials into osteogenic or adipogenic lineage between two groups, alizarin red S and oil red O staining were performed at 14 days after induction of differentiation in vitro. RESULTS: Flow cytometry results showed that early passage tendon-derived cells expressed CD44 in 99.13%, CD90 in 56.51%, and CD105 in 86.19%. In the CFU-F assay, CD90+ or CD105+ cells isolated with MACS showed larger colony formation in size than cells isolated using the colony picking method. We also observed that CD90+ or CD105+ cells were constantly differentiated into both osteogenic and adipogenic lineages in cells from all donors, whereas cells isolated using the colony picking method were heterogeneous in differentiation potentials to the osteogenic and adipogenic lineages. CONCLUSION: CD90+ or CD105+ cells isolated using MACS showed superior MSC characteristics in the self-renewal and multi-differentiation capacities compared with cells isolated using the colony picking method.
