前列腺原代基质细胞培养方法的研究

Primary Culture of Benign Prostatic Hyperplasia Stromal Cell

目的建立前列腺原代基质细胞(PS)培养的方法,并研究年龄与胶原打底对于PS的影响。方法对老年男性前列腺组织分别进行酶消化法、组织块法、酶消化法+组织块法PS培养。通过免疫细胞化学染色对PS进行鉴定。结果组织块法成功培养PS。酶消化法中PS细胞接种后3 d贴壁率仅6.4%,细胞2周已经老化。酶消化法+组织块法培养细胞无法贴壁。胶原打底组细胞贴壁率与生长状态优于普通瓶底组,供者年龄<60岁的PS贴壁率与生长状态优于供者年龄≥60岁的PS。结论组织块法简单易行,适于行前列腺PS培养。胶原打底与供者年龄<60岁的原代细胞培养更适合行PS培养。

Objective To investigate the primary culture technique in vitro,and to evaluate the influence of the prostate donor age and the collagen coating on the primary prostatic cell culture.Methods Three different methods（enzymic digestive method,tissue culture and enzymic digestive method combined with tissue culture） were investigated in the primary prostatic cell culture,the stromal cell were identified by immunocytochemistry.Results Compared with the methods of digestion（either with or without tissue culture）,tissue culture possessed better cellular adherence and proliferative ability.Thin coating of collagen on culture flask could improve the adherence of cell and promote cell proliferation.Prostate stromal cells from donors aged ≥60 years old grew slower than those from donors aged 〈60 years old.Conclusion Tissue culture is a simple,convenience method for the primary benign prostatic hyperplasia（BPH） stromal cell culture.Thin coating of collagen in culture flask could improve the primary culture,and prostate stromal cells from donors aged 〈60 years old are more suitable for stromal cell culture.