Cell Culture of Human Degenerative Intervertebral Disc
摘要
BACKGROUND: There are no reports about adult degenerative disc cell culture model currently. This establishment of disc cell culture is the cellular basis of intervertebral disc tissue engineering. OBJECTIVE: To establish cell culture models from human degenerative intervertebral disc, and to settle the cytology base of disc tissue engineering. DESIGN AND SETTING: The controlled observational experiment with regard to cytology was performed in Shandong Institute of Orthopaedics and Traumatology. MATERIALS: Five degenerative intervertebral disc patients were recruited from Department of Orthopaedics, Affiliated Hospital of Qingdao University Medical College, including one case in L2-3, three cases in L4-5 and one case in L5-S1. METHODS: Five samples were taken from the operational disectomy for lumbar disc herniation, and these samples were cultured using enzymatic digestion cell culture and tissue cell culture method differently. All the samples were cultured with HAMF12 medium with the addition of 10% and 20% fetal bovine serum. The morphology of the cultured cell was observed with Giemsa staining and transmission electron microscopy respectively. MAIN OUTCOME MEASURES: ① Cellular growth; ② Morphology of the cultured cells; ③ Ultrastructure of the cultured cells. RESULTS: By means of enzymatic digestion cell culture and tissue cell culture method, numerous degenerative lumbar disc cells were obtained and successfully subcultured. The cell secretions around the cultured cells may influence the growth of these cells. Destroying the secretions, the cultured cells could be highly proliferated. The degenerative intervertebral disc cells were proliferated vigorously in HAMF12 medium with 20% fetal bovine serum compared with that with 10% fetal bovine serum. The notochordal cell was observed in all the specimens. CONCLUSION: The adult degenerative disc can be used to obtain the required cell for tissue engineering.