Objective To study the relationship of different ages with harvesting of ligamentum flavum and cell growth rate in primary culture, so as to improve the success rate of human ligamentum flavum cells(LFCs) culture in vitro. Methods A total of 21 surgical ligamentum flavum specimens from patients with intervertebral disc herniation and lumbar spinal stenosis were primary cultured with tissue block enzymolytic method. Cell outgrowth, morphology and growth status were observed under an inverted phase microscope. Expression of vimentin and type Ⅰ collagen was detected in the cells of third passage by immunofluorescence staining. The correlation between age and cell growth rate was analyzed. Results Cell morphology and growth status were well under inverted phase microscope and immunofluorescence staining showed vimentin and collagen type Ⅰ expression in the all LFCs. The cell growth rate of primary culture was negatively correlated with the age of patients from whom ligamentum flavum had been collected(r=-0.618, P<0.05). Cell growth rate was high when the patients were young. Conclusion Ligamentun flavum specimens should be taken from young subjects to ensure the success rate of primary culture of LFCs.