The Experimental Study Of Human Annulus Fibrosus Cell And Bone Marrow Mesenchymal Stem Cells Co-culture By Transwell

Background：Intervertebral Disc degeneration often cause neck shoulder pain and lowback pain. The surgical treatment of the commonly used method is provides good short-termcurative effect, but the change of the spine surgery biomechanics, adjacent to the intervertebraldisc further degeneration. So, looking for Intervertebral disc degeneration in early interventionand maintain of spine biomechanics of tissue engineering repair methods, become the focus ofthe present study. Bone marrow mesenchymal stem cells (BMSCs) is to become the discdegeneration repair stem cell therapy hot spot. annulus fibrosus (AF) is an important componentof the disc structure, Its structure and function in the integrity of the disc degeneration in theprevention and control of great significance.Objective: Discusses the human annulus fibrosus (AF) cell and the bone marrowmesenchymal stem cells (BMSCs) co-culture in Transwell on AF cells cell proliferation andextracellular matrix expression, and the BMSCs whether it has the ability to AF celldifferentiation.Methods: For respectively between five the following ages of40years old spinal surgerypatient AF cells and BMSCs separation,training.The third passage of BMSCs and AF cell wereharvested and placed into Transwell co-culture system at a ratio of1:1, BMSCs were placed inthe bottom and AF cell in the upper. Three groups were designed as follows，BMSCs group andAF cell group as control and co-cultured group termed as test group.After three week, the shapechanges of these cell were observed under phase contrast microscopy; Use HE dyeing, typeⅠcollagen immunohistochemical staining and flow cytometric to two kinds of cells identificationrespectively and the changes of mRNA of typeⅠ collagen、Sox9and proteoglycan of these threetypes of cells were detected by RT-PCR respectively.Result: BMSCs and AF cell were identified. Under phase contrast microscopy, AF celland BMSCs in co-cultured group for cell proliferation quantity and speed are higher than thosein control group. while the mRNA of type Ⅰ collagen、Sox9and proteoglycan of AF cell and BMSCs in co-cultured group increased higher than those in control groups （P<0.05）.Conclusion: Through the BMMSCs and AF cells Transwell co-culture, BMSCs canpromote AF cell proliferation and extracellular matrix synthesis and the BMSCs has the ability toAF cell differentiation.