Chinese Journal of Tissue Engineering Research ›› 2013, Vol. 17 ›› Issue (7): 1168-1173.doi: 10.3969/j.issn.2095-4344.2013.07.006

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Adult degenerative nucleus pulpous cells cultured by a microcarrier-based stirred culture system

Ning Bin1, Ding Yuan-jing1, Gong Wei-ming1, Liu Hai-fei2, Liu Yong2, Wang De-chun2, Hu You-gu2   

  1. 1 Department of Spinal Surgery, Jinan Central Hospital Affiliated to Shandong Universtiy, Jinan 250013, Shandong Province, China
    2 Orthopedic Research Institute of Shandong Province, Qingdao 277018, Shandong Province, China
  • Received:2012-07-28 Revised:2012-09-11 Online:2013-02-12 Published:2013-02-12
  • Contact: Hu You-gu, Professor, Master’s supervisor, Orthopedic Research Institute of Shandong Province, Qingdao 277018, Shandong Province, China huyougu@hotmail.com
  • About author:Ning Bin☆, Doctor, Attending physician, Department of Spinal Surgery, Jinan Central Hospital Affiliated to Shandong Universtiy, Jinan 250013, Shandong Province, China ningbin@mail.sdu.edu.cn

Abstract:

BACKGROUND: Rotating bioreactor is a microcarrier-based stirred culture system that is currently used more often.
OBJECTIVE: To culture the adult degenerative nucleus pulposus cells in vitro by a microcarrier-based stirred culture system, and then to amplify and identify the cells.
METHODS: Nucleus pulposus specimens taken from patients with disc degenerative diseases were randomly divided into monolayer culture group and micro-carrier culture group. The cell morphology was observed by hematoxylin-eosin staining and Gimsa staining, the microstructure was observed under an electron microscope, cell viability was detected by MTT method.
RESULTS AND CONCLUSION: The morphology of degenerative nucleus pulpous cells was identified by inverted phase contrast microscope, and the regression was showed by a transmission electron microscope. The microcarrier cultured cells were observed by an inverted phase contrast microscope and scanning electron microscope showing stereoscopic shape. In the logarithmic growth phase, the growth activity of microcarrier culture cells was significantly higher than that of the cells in monolayer culture. Microcarrier-based stirred culture system can stabilize the cell phenotype, and significantly stimulate the cellular activity of nucleus pulposus cells in the logarithmic growth phase.

Key words: tissue construction, cartilage tissue construction, nucleus pulpous cells, degeneration, microcarriers, cell culture, cell identification, three-dimensional culture, mondayer culture, the National Natural Science Foundation of China, tissue construction photographs-containing paper

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