Objective To establ ish a two-dimensional biological printing technique of hBMSCs so as to control the cell transfer process and keep cell viabil ity after printing. Methods Bone marrow (5 mL) was obtained from healthy volunteer. The hBMSCs were regularly subcultured to harvest cells at passage 2, which were adjusted to the single cell suspension
at a density of 1 × 106/mL. The experiment was divided into 3 groups: printing group 1 in which cells underwent propidium iodide (PI) fluorescent label ing, then were transferred by rapid prototype biological printer (interval in x-axis 300 μm, interval in y-axis 1 500 μm), and laser scanning confocal microscope was appl ied to observe cell fluorescence; printing group 2 in which cells received no PI label ing and were cultured for 2 hours after transfer, Live/Dead viabil ity Kit was adopted to detect cell viabil ity and laser scanning confocal microscope was appl ied to observe cell fluorescence; half of the cells in printing group receiving no Live/Dead viabil ity Kit detection were cultured for 7 days, then inverted microscope was used to observe cell morphology, routine culture was conducted after the adherence of cells, the growth condition of cells was observed dynamically; control group in which steps were the same as the printing group 2 except that cell suspension received no printing. Results Laser scanning confocal microscope observation on the cells in printing group 1 revealed the “cell ink droplets” were distributed regularly and evenly in the two-dimensional layer and each contained 15-35 cells, meeting the requirement of designing two-dimensional cell printing. The cells in printing group 2 went through cell viabil ity test, laser scanning confocal microscope observation showed the fluorescence of cells 30 minutes after cell incubation. There was no significant difference between the control group and the printing groups in terms of cell viabil ity. The printed cells presented normal adherence, good morphology and good growth state 7 days after routine culture. Conclusion Biological printing technique can real ize the oriented, quantificational and regular
distribution of hBMSCs in the two-dimensional plane and lays the foundation for the construction of three-dimensional cell
printing or even organ printing system.
Citation： MA Sunxiang,CHAI Gang,LIU Qihai,HU Qingxi,ZHOU Guangdong,CUI Lei. PRELIMINARY STUDY ON BIOLOGICAL PRINTING OF hBMSCs. Chinese Journal of Reparative and Reconstructive Surgery, 2009, 23(4): 497-500. doi: Copy