Leishmania donovani: use of two different, commercially available, chemically defined media for the continuous in vitro cultivation of promastigotes.
Leishmania donovani: use of two different, commercially available, chemically defined media for the continuous in vitro cultivation of promastigotes.
复制标题
杜氏利什曼原虫：使用两种不同的市售化学成分确定的培养基来连续体外培养前鞭毛体。
DOI：
10.1016/0014-4894(91)90112-a
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发表时间：
1991
影响因子：
2.1
通讯作者：
D. Dwyer
中科院分区：
文献类型：
--
作者：
C. McCarthy;P. Bates;D. Dwyer
培养基 199 和 RPMI-1640 用作 2 种限定培养基制剂的基础。 M199+含有用侧翼盐缓冲的培养基199和25msmallcap~M Hepes，并补充有叶酸、血红素、青霉素、链霉素和基础培养基Eagle维生素。 RPMI+由RPMI-1640和25msmallcap~M Hepes和用与培养基199+相同的成分强化的smallcap~L-谷氨酰胺和腺苷组成。实验培养物从 L 的克隆系开始。多诺瓦尼前鞭毛体 (MHOM/SD/00/1S-2D)。在 M199+ 和对照 M199、10% FBS 中生长的寄生虫的等效世代时间为 8.9±0.1 小时，而在 RPMI+ 中生长的寄生虫的平均倍增时间为 15.8±0.05 小时。一旦适应了培养基，100 多次连续传代的平均生成时间就保持稳定。与所使用的培养基无关，前鞭毛体在对数晚期/稳定期早期达到了 3-4×107 个细胞/ml 的等效细胞密度。前鞭毛体连续繁殖一年多。L.墨西哥(MNYC/BZ/62/M379)，L。亚马逊 (MHOM/BR/73/M2264),L. Major (MHOM/IL/80/Friedlin) 和 Crithidiae lucilia 也已连续培养。 附加摘要：本文报道了在市售的、限定的、无大分子的培养基中在未限定的温度 [大概 26 °C] 下成功长期培养杜氏利什曼原虫前鞭毛体的克隆。论文中给出了完整的细节；使用的2种培养基是(i)含有Hanks盐的培养基199，用HEPES缓冲并补充有血红素和叶酸(加抗生素)，和(ii)具有HEPES和小帽~L-谷氨酰胺的RPMI-1640，补充有血红素、叶酸和腺苷(加抗生素)。前者的生长更快（倍增时间约为 9 小时，而倍增时间约为 16 小时），但达到了 3-4 × 107 前鞭毛体/ml 的等效密度。 mexicana、L. amazonensis、L. Major 和 Crithidia luciliae 也已在这些确定的培养基中生长。贝克
Medium 199 and RPMI-1640 were used as the bases for 2 defined media preparations. M199+ contained Medium 199 buffered with flanks' salts and 25 msmallcap˜M Hepes, and was supplemented with folic acid, haemin, penicillin, streptomycin and basal medium Eagle vitamins. RPMI+ consisted of RPMI-1640 with 25 msmallcap˜M Hepes and smallcap˜L-glutamine fortified with the same components as for Medium 199+, and adenosine. Experimental cultures were initiated from a cloned line ofL. donovanipromastigotes (MHOM/SD/00/1S-2D). Parasites grown in M199+, and in the control M199, 10% FBS had equivalent generation times of 8.9±0.1 h, whereas those grown in RPMI+ had a mean doubling time of 15.8±0.05 h. The mean generation time remained stable for over 100 serial passages once they were adapted to the media. Independent of the media used, promastigotes attained equivalent cell densities of 3-4×107cells/ml at late log/early stationary phase. Promastigotes were serially propagated for over a year.L. mexicana(MNYC/BZ/62/M379),L. amazonensis(MHOM/BR/73/M2264),L. major(MHOM/IL/80/Friedlin) andCrithidiae luciliahave also been serially cultivated.ADDITIONAL ABSTRACT:This paper reports the successful long-term cultivation of a clone ofLeishmania donovanias promastigotes in commercially available, defined, macromolecule-free culture media at an undefined temperature [presumably 26 °C]. Full details are given in the paper; the 2 media used were (i) Medium 199 with Hanks' salts buffered with HEPES and supplemented with haemin and folic acid (plus antibiotics), and (ii) RPMI-1640 with HEPES and smallcap˜L-glutamine, supplemented with haemin, folic acid and adenosine (plus antibiotics). Growth was more rapid in the former (doubling time about 9 h compared with about 16 h), but equivalent densities of 3-4 × 107promastigotes/ml were achieved.Isolates ofL. mexicana, L. amazonensis, L. majorandCrithidia luciliaehave also been grown in these defined media.newline˜J.R. Baker