J Korean Assoc Maxillofac Plast Reconstr Surg.
2005 Jan;27(1):1-8.
Problems in osteogenic differentiation of rat bone marrow stromal cells
- Affiliations
-
- 1Department of Oral & Maxillofacial Surgery, College of Dentistry, Seoul National University, Korea. sjhwang@snu.ac.kr
- 2Dental Research Institute, Seoul National University, Korea.
- 3Department of Biomedical Engineering, College of Medicine, Brain Korea 21 Program for Biomedical Science, Korea Artificial Organ Center, Korea University, Korea.
- 4Chemical Engineering Department, Seoul National University of Technology, Korea.
- 5Laboratory for Osteology, Department of Craniomaxillofacial Surgery, University Hospital Zurich, Switzerland.
Abstract
- This study was aimed to characterize osteogenic potential of rat bone marrow stromal cells (BMSC) isolated with standard flushing method and investigate the plasticity of transdifferentiation between osteoblastic and adipocytic lineage of cultured BMSC. Unlike aspiration method in human, rat bone marrow was extracted by means of irrigation with culture media that elevates the possibility of co-extraction of committed osteoprogenitor, or preosteoblast or other progenitor cells of several types present inside bone marrow. The cultured stromal cells showed high ALP activity which is representative marker of osteoblast without any treatment. Osteogenic inducers such as Dex and BMP-2 were examined for the evaluation of their effect on osteogenic and adipocytic differentiation of stromal cells, because they function as osteoinductive agent in stromal cells, but simultaneously induce adipogenic differentiation. Osteogenic differentiation was evaluated by measuring alkaline phosphatase activity or mRNA expression of osteoblast markers such as osteopontin, bone sialoprotein, collagen type I and CbfaI, and in vitro matrix mineralization by von Kossa staining. Oil red staining method was used to detect adipocyte and adipocytic marker, aP2 and PPARgamma2 expression was examined using RT-PCR. It can be supposed that irrigation procedure resulted in high portion of already differentiation-committed osteoprogenitor cell showing elevated ALP activity and strong mineralization only under the supplement of 100microM ascorbic 2-phosphate and 10mM beta-glycerophosphate without any treatment of osteogenic inducers such as Dex and BMP-2. Dex and BMP-2 seemed to transdifferentiate osteoprogenitor cells having high ALP activity into adipocytes temporarily, but continuous treatment redifferentiated into osteoblast and developed in vitro matrix mineralization. This property must be considered either in tissue eng ineering for bone regeneration, or in research of characterization of osteogenic differentiation, with rat BMSC isolated by the standard irrigation method.
Keyword
MeSH Terms
-
Adipocytes
Alkaline Phosphatase
Animals
Bone Marrow*
Bone Regeneration
Collagen Type I
Culture Media
Flushing
Humans
Integrin-Binding Sialoprotein
Mesenchymal Stromal Cells*
Osteoblasts
Osteopontin
Plastics
PPAR gamma
Rats*
RNA, Messenger
Stem Cells
Stromal Cells
Alkaline Phosphatase
Collagen Type I
Culture Media
Integrin-Binding Sialoprotein
Osteopontin
PPAR gamma
Plastics
RNA, Messenger
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