To find a new source of seed cells for constructing tissue-engineered intervertebral disc,nucleus pulposus(NP) cells and mesenchymal stem cells(MSCs) were isolated from New Zealand white rabbits.The nucleus pulposus cells population was fluorescence-laelled and co-cultured with MSCs with or without direct contact.Morphological changes were observed every 12 h.Semi-quantitaive reverse transcriptase-polymerase chain reaction was performed to assess the expression levels of Sox-9,aggreacan and type Ⅱ collagen every 24 h after the co-culture.MSCs treated with direct contact rounded up and presented a ring-like appearance.The expression of marker genes was significantly increased when cells were co-cultured with direct contact for 24 h.No significant change was found after coculture without direct contact.Co-culture of NP cells and MSCs with direct contact is a reliable method for generating large amount of NP cells used for cell-based tissue engineering therapy.
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