Construction of in vitro models of human bone marrow-derived mesenchymal stem cells differentiated into nucleus pulposus-like cells in rabbits

Quanxiu LI; Bohua CHEN; Yong LIU; Ling XU

ABSTRACT

BACKGROUND:

There is notan effective method for inducing bone marrow-derived mesenchymal stem cells (BMSCs) differentiatied in to intervertebral disc cells.

OBJECTIVE:

To establish a model of human BMSCs (hBMSCs) differentiated in to nucleus pulposus-like cells in vitro.DESIGN,TIME AND

SETTING:

A single sample contrast.The experiment was performed at the Orthopaedics Institute and Central Laboratory,the Affiliated Hospital of Qingdao University Medical College,from August 2008 to March 2009.MATERIALSTwo New Zealand white rabbits with 8-week-old were used to culture nucleus pulposus cells.Human nucleus pulposus was obtained from a aged 16 years patient with T_(12)L_1,L_(1.2) congenital scoliosis.The informed consent was obtained from his patients,hBMSCs were purchased from Salye Biological Science and Technology Co.,Ltd.(Guangzhou,China).

METHODS:

hBMSCs and nucleus pulposus cells of rabbit were co-cultured in new-style culture model (in 6-well plates and inserts with track-etched membrane having 0.4 μm pores without arms) for 7 days,conventionally laminar culture,solitary hBMSCs and nucleus pulpesus cells culture were set up as controls at same time.MAIN OUTCOME

MEASURES:

The expressions of collagen Ⅱ and aggrecan mRNA were detected by RT-PCR and Western blot.

RESULTS:

hBMSCs in the conventionally laminar and single hBMSCs groups were negative to collagen Ⅱ and aggrecan.The collagen Ⅱ and aggrecan expression of hBMSCs were upregulated evidently in new-style culture group,which was similar to nucleus pulposus cells that from degenerate intervertebral disc of human,but these expression did not appear in conventionally laminar culture and solitary hBMSCs culture (P ＜ 0.01).The difference among new-style culture group and other groups had no significance (P ＞ 0.05).

CONCLUSION:

A new-style model of hBMSCs differentiation to nucleus pulposus-like cells is constructed in vitro,which can highly express collagen Ⅱ and aggrecan and provide a foundation for intervertebral disc degeneration cellular treatment.