悬浮培养型 BHK -21 细胞的生长规律和不同接种密度培养效果的研究

为探索悬浮培养型BHK-21细胞的生长规律和最佳接种密度，以3．0×10^5个／mL、4．0×10^5个／mL、5．0×10^5个／mL、6．0×10^5个/mL和7．0×10^5个／mL等5个接种密度培养了悬浮培养型BHK-21细胞，绘制了细胞生长曲线和活力变化曲线．结果显示，悬浮培养型BHK-21细胞生长曲线呈“S”型，接种培养前24h细胞处于适应期，生长曲线上密度增长不大，期后细胞进入对数增长期，生长密度呈指数增长．以3．0×10^5个／mL和4．0×10^5个／mL接种培养的在120h达到最大增殖密度，分别为4．65×100个／mL和5．59×10^6个／mL，倍增时间分别为26．9h和26．7h；以5．0×10^6个／mL、6．0×10^6个/mL和7．0×10^6个／mL接种培养的在96h就达到最大增殖密度，分别为5．14×10^6个／mL、5．36×10^6个／mL和5．1×10^6个／mL，倍增时间分别为22．4h、24．1h和25．2h．各组在培养的96h以前细胞活力均在90％以上且变化较小，120h和144h时细胞活力开始下降，且接种密度越高活力下降越快．该细胞以4．0×10^5个／mL、5．0×10^5个／mL、6．0×10^5个／mL接种培养能达到较高培养密度，细胞生长快，且细胞峰值持续时间长．

Growth Rhythm and Effects in Different Inoculation Density of BHK- 21 Suspension Cell Culturing

To explore the growth rhythm and optimum inoculation density of BHK- 21 suspension cell, five cell inoculation densities of 3.0 ×10^5cells/mL, 4.0 ×10^5 cells/mL, 5.0 ×10^5cells/mL, 6.0 ×10^5 cells/ mL and 7.0×10^5 cells/mL have been tested in the experiments. The cell growth and vitality curves were drawn. The results showed that the growth curve of BHK - 21 suspension cell was ＂S＂ style. 24 h after cell inoculation was adaptive phase, which the cell density increased very small, and then the logarithmic phase was come in, during which the cell density increased exponentially. The maximum proliferation concentrations of cell inoculation densities for 3.0 ×10^5 cells/ml and 4.0 ×10^5 cells/mL were 4.65 ×10^6 cells/mL and 5.59 ×10^6 cells/mL in 120 h, respectively. The maximum proliferation concentrations of cell inoculation densities for 5.0 ×10^5cells/mL, 6.0 ×10^5 cells/mL and 7.0 ×10^5cells/mL were 5.14 ×10^6 cells/mL, 5.36 ×10^6 cells/mL and 5.1 ×10^6 cells/mL in 96 h, respectively. The population doubling times （PDT） were respectively 22.4 h, 24.1 h and 25.2 h. The cell vitalities in all groups were above 90 % before 96 h and changed very small. The cell vitalities began to decline after 120 h and 144 h. The BHK - 21 suspension cell in inoculation densities of 4.0 ×10^5 cells/mL, 5.0 ×10^5 cells/mL and 6.0 ×10^5 cells/mL can achieve higher cell density, the cell growth was fast, and cell peak duration was long.