Biotin and choline replace the growth requirement of Madin-Darby canine kidney cells for high-density lipoproteins.
Biotin and choline replace the growth requirement of Madin-Darby canine kidney cells for high-density lipoproteins.
复制标题
生物素和胆碱替代了 Madin-Darby 犬肾细胞对高密度脂蛋白的生长需求。
DOI：
10.1002/jcp.1041240116
复制
发表时间：
1985
影响因子：
5.6
通讯作者：
Gospodarowicz,D
中科院分区：
文献类型：
--
作者：
Cohen,DC;Gospodarowicz,D
MDCK细胞保存在细胞外基质（ECM）包被的培养皿中，并暴露于添加转铁蛋白和高密度脂蛋白（hdl）或磷脂酰胆碱（PC）脂质体的Dulbecco改良Eagle培养基（DME）中，其生长速度和最终细胞密度与暴露于添加血清DME的培养物相似。当MDCK细胞暴露于由DME和F12培养基（D/F）的混合物（1:1）组成的培养基中时，仅添加转铁蛋白（10 μg/ml）即可支持细胞生长，并且不再需要hdl或PC脂质体的存在。MDCK细胞暴露于添加转铁蛋白的D/F培养基中，在完全没有血清的情况下可以传代50代以上。在缺乏hdl或PC脂质体的情况下，支持生长的F12成分是生物素（二甲醚中不存在）和胆碱（二甲醚中存在浓度不足）。在DME中添加转铁蛋白、生物素（3.6 ng/ml）和胆碱（10 μg/ml）可使MDCK细胞生长最佳，并可连续繁殖50代以上。因此，MDCK细胞对hdl或PC脂质体的生长需求可以由足够浓度的生物素和胆碱代替。二甲醚/F12培养基的组合比单用二甲醚在支持细胞生长方面更有效，这一事实被广泛观察到，部分原因可能是二甲醚中缺乏生物素和次理想的胆碱浓度。
MDCK cells maintained on extracellular matrix (ECM)‐coated dishes and exposed to Dulbecco's modified Eagle's medium (DME) supplemented with transferrin and either high‐density lipoproteins (HDLs) or phosphatidyl choline (PC) liposomes have a growth rate and final cell density similar to those of cultures exposed to serum‐supplemented DME. When MDCK cells are exposed to a medium consisting of a mixture (1:1) of DME and F12 medium (D/F), the addition of transferrin (10 μg/ml) alone supports cell growth and the presence of HDLs or PC liposomes is no longer required. MDCK cells exposed to D/F medium supplemented with transferrin can be passaged for more than 50 generations in total absence of serum. The F12 components that support growth in the absence of HDLs or PC liposomes are biotin (which is absent in DME) and choline (which is present in insufficient concentration in DME). Supplementation of DME with transferrin, biotin (3.6 ng/ml), and choline (10 μg/ml) allows optimal growth of MDCK cells and permits serial propagation through more than 50 generations. The growth requirement of MDCK cells for HDLs or PC liposomes can therefore be replaced by adequate concentrations of biotin and choline. The widely observed fact that a combination of DME/F12 medium is more effective than DME alone in supporting cell growth may be due in part to the lack of biotin and suboptimal choline concentration in DME.