title: [The changes of extracellular matrix in adult degenerative nucleus pulposus cells with stiring microcarrier culture system in vitro].
abstract: OBJECTIVE
  To evaluate the biological effect on the synthesis of the extracellular matrix (ECM) in the cultivation of adult degenerative nucleus pulposus cells using the stiring microcarrier system in vitro.
  
  
  METHODS
  Thirty-four specimens were collected after intervertebral fusion operations of the patients with intervertebral disc herniation diseases from September 2005 to May 2009. The specimens were then randomly allocated into 2 groups for in vitro cultivation: monolayer culture group and microcarrier culture group. On the exponential phase, SP-ABC immunohistochemical staining and Western blot quantitative analysis were conducted in the two groups to detect the collagen type I and II. Proteoglycan contents of two groups in different growth phases were detected with (35)S-sulfate incorporation assay.
  
  
  RESULTS
  The expressions of collagen type I and II in microcarrier culture group were significantly higher than those in monolayer culture group: SP-ABC immunohistochemical staining (collagen type I: 32.5 ± 4.4 vs. 15.2 ± 1.2, t = 2.871, P < 0.01; collagen type II: 43.6 ± 4.1 vs. 23.1 ± 2.2, t = 2.375, P < 0.05); Western blot quantitative analysis (collagen type I: 0.62 ± 0.08 vs. 0.50 ± 0.06, t = 3.327, P < 0.01; collagen type II: 1.46 ± 0.08 vs. 0.86 ± 0.04, t = 2.453, P < 0.05). Nucleus pulposus cells cultivated in stiring microcarrier system showed significantly increased proteoglycan synthesis than monolayer culture group does on both exponential phase and stationary phase (exponential phase: 34 821 ± 312 vs. 21 046 ± 673, t = 2.134, P < 0.05; stationary phase: 45 134 ± 175 vs. 32 193 ± 713, t = 2.801, P < 0.01).
  
  
  CONCLUSIONS
  The expression of collagen type I, II and proteoglycan of adult degenerative nucleus pulposus cells are positive regulated by the stiring microcarrier system, which can be used in the mass amplification of the adult degenerative nucleus pulposus cells.
authors: Department of Orthopaedic, Jinan Central Hospital Affiliated to Shandong University, Jinan 250013, China.
authors: Bin
authors: B
authors: Ning
authors: Hai-fei
authors: HF
authors: Liu
authors: Wei-ming
authors: WM
authors: Gong
authors: Kai
authors: K
authors: Zhao
authors: Hong-xia
authors: HX
authors: DU
authors: Yong
authors: Y
authors: Liu
authors: De-chun
authors: DC
authors: Wang
authors: You-gu
authors: YG
authors: Hu
mesh_terms: D000328:Adult; D000368:Aged; D018929:Cell Culture Techniques; D003094:Collagen / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism*; D005260:Female; D006801:Humans; D007403:Intervertebral Disc / Q000166:cytology*; D008297:Male; D008875:Middle Aged; D011509:Proteoglycans / Q000378:metabolism; D011897:Random Allocation; D055815:Young Adult
chemical_list: D011509:Proteoglycans; D003094:Collagen