Human placenta-derived mesenchymal stem cells (PMSCs) are a kind of important seed cells for cell therapy, regenerative medicine, as well as immunoregulatory treatment. The study on the culture surface is a key topic to optimize PMSCs culture system.OBJECTIVE:
To observe the biological effects of polystyrene culture surfaces modified by sulfonation, sub-atmospheric pressure plasma technology or poly-D-lysine (PDL)-coated technology on PMSCs cultured in vitro.DESIGN, TIME AND SETTING:
An in vitro contrast experiment was performed at Laboratory of Bioplastic, Beijing Transglobal Bio-medical Technological Co., Ltd. from March 2007 to October 2008.MATERIALS:
Polystyrene culture dish (3.5 cm2) and 24-well culture plate were provided by Beijing Transglobal Bio-medical Technological Co., Ltd.; poly-D-lysine was provided by Sigma, USA; normal human placenta was provided by Beijing People's Hospital and the informed consent was obtained from puerpera.METHODS:
Polystyrene culture surfaces were modified by sulfonation, sub-atmospheric plasma technology, and PDL-coated technology to increase their surface tension. Human placenta lobule nucleated cells were separated, and inoculated into above modified culture surfaces. PMSCs colonies were harvested by adherence method and then passaged.MAIN OUTCOME MEASURES:
Polystyrene surface tension, PMSCs primary culture cloning rates, adherent rates after inoculated, and growth curves.RESULTS:
The polystyrene surface tension after modified by different methods was as follows: plasma group > PDL group > sulfonation group > control group; the greater the surface tension was, the greater the primary culture cloning rate and subculture adherent rate of human PMSCs were. The PDL-coated surface was more suitable for human PMSCs primary culture. Increasing surface tension could effectively shorten PMSCs recovery period after inoculation.CONCLUSION:
Whereas the significant increase of PDL-coated costs and the possibility of cell contamination, we recommend that PMSCs primary culture was performed using PDL-coated surface, and PMSCs passage culture was performed using culture surface treated by plasma modification.