Osteogenic Potential of Human Mesenchymal Stem Cells During Serial Subculture.
- Author:
Hyun Jin SUN
1
;
Yon Rak CHOI
;
Soo Bong HAHN
;
Jin Woo LEE
Author Information
1. Brain Korea 21 project for medical science, Department of Orthopaedic Surgery, Yonsei University College of Medicine, Seoul, Korea. ljwos@yumc.yonsei.ac.kr
- Publication Type:In Vitro ; Original Article
- Keywords:
Mesenchymal stem cell;
Subculture;
Osteoblast;
Differentiation
- MeSH:
Alkaline Phosphatase;
Coloring Agents;
Humans*;
Mesenchymal Stromal Cells*;
Osteoblasts;
Phenotype;
RNA, Messenger
- From:Journal of Korean Orthopaedic Research Society
2004;7(2):99-107
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
PURPOSE: The purpose of this study was to evaluate the osteogenic differentiation potential of human mesenchymal stem cells during serial subculture. MATERIALS AND METHODS: Human bone marrow-derived MSCs were serially subcultured and then maintained in basal or osteogenic medium for 14 days. Then we performed FAC analysis, RT-PCR, alkaline phosphatase activity and stains. RESULTS: Human MSCs had different morphologies, immunophenotypes, and growth rates that were correlated with the length of serial subculture. The phenotype changed from small spindle-shaped cells at passage 1 into large cuboidal or flattened cells at passage 7. The osteogenic capacity of human MSCs decreased during serial subculture. Using RT-PCR, the mRNA levels of bone-specific genes, such as cbfa1/runx2 and osteocal-cin, decreased with increasing passage number. Strong positive staining was observed for ALP and Alizarin reds in osteogenic medium on day 14, but declined significantly with increasing passage number. CONCLUSION: We have shown that osteogenic potential of human MSCs decreased during serial subculture. This result can provide the helpful information to decide the timing of human MSC transplantation during in vitro culture expansion for treatment of bone defects and so on.
