Characteristics of bone marrow-derived mesenchymal stem cells isolated in vitro with osteogenic potential under induced condition

Jisheng XIE; Zansong HUANG; Yujin TANG; Hailing HUANG; Minan LU; Qiongzhi ZHAO

ABSTRACT

BACKGROUND:

Based on the multi-directional differentiation potential,the combined application of proper biological materials and growth factor can repair tissue defect.

OBJECTIVE:

To observe the growth characteristics of rabbit bone marrow-derived mesenchymal stem cells and their osteogenic potential under the induced condition.

DESIGN:

A single sample experiment.

SETTING:

The Department of Histology and Embryology of Youjiang Medical College for Nationalities; the Affiliated Hospital of Youjiang Medical College for Nationalities.MATERIALS This experiment was conducted at the Department of Histology and Embryology of Youjiang Medical College for Nationalities in June 2004. Male new Zealand rabbits, with a body mass of 2.0 to 2.5 kg,were provided.

METHODS:

Rabbit bone marrow tissue was extracted. After gradient centrifugation, adherent cells were kept for cell passage. Culture medium was used for inducing culture of osteoblast when the passage was stable (50 mL/L of Dulbecco modified culture medium of bovine serum containing 10 mmol/L of β-sodium glycerophosphate, 10 nmol/L of dexamethasone, 50 mg/L of vitamine C). The culture medium was changed every other day. Osteogenic characteristics of the bone marrow-derived mensenchymal stem cells were observed under an inverted microscope after alkaline phosphatase immunohistological staining, osteonectin and osteopontin immunocytochemical staining.MAIN OUTCOME

MEASURES:

① Morphological observation of bone marrow-derived mesenchymal stem cells; ② Osteogenic characteristics of bone marrow-derived mesenchymal stem cells.

RESULTS:

① Morphological observation of bone marrow-derived mensenchymal stem cells Primary rabbit bone marrow-derived mesenchymal stem cells could cover the Petri dish fully in 7 or8 days and could passage stably. Generative cells began to passage in 5 or 6 days. ② Osteogenic characteristics of bone marrow-derived mensenchymal stem cells Alkaline phosphatase immunohistochemical staining showed a lot of brown particles in the kytoplasm, but no colored cells were found in the control group. Osteonectin and osteopontin immunohistochemical detection found that the nucleus was light blue and a lot of brown particles appeared in the kytoplasm, presenting obvious strong positive, but no brown particles appeared in the control group.

CONCLUSION:

After combined induced culture with dexamethasone, vitamine C and β-sodiumglycerophosphate, rabbit bone marrow mensenchymal stem cells showed the morphological and biological characteristics of osteoblasts with osteoblastic activity, and were able to supply autologous seed cells for constructing in vitro tissue engineered bone in a short period of time.