In vitro cloning of tumor stem cells in semi-solid media containing agar and agarose.
In vitro cloning of tumor stem cells in semi-solid media containing agar and agarose.
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在含有琼脂和琼脂糖的半固体培养基中体外克隆肿瘤干细胞。
DOI：
10.1007/bf00199774
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
Kushner,H
中科院分区：
文献类型：
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作者：
Bagasra,O;Berman,W;Hashemi,S;Howeedy,A;Kushner,H
通过比较三种小鼠畸胎癌衍生细胞系和一种人畸胎癌衍生细胞系在含有琼脂或琼脂糖的半固体培养基中的生长，研究了肿瘤干细胞集落的体外形成。我们表明，琼脂糖的使用浓度应高于琼脂才能获得可比较的结果。与琼脂相比，在更广泛的浓度范围（SeaPrep 为 1%–4%，SeaPlaque 琼脂糖为 0.5%–2%）的琼脂糖中获得最大集落数，这使得肿瘤细胞仅在较窄的浓度范围（0.3%–0.5%）内进行贴壁独立生长。总体而言，含有琼脂糖的培养基的制备比含有琼脂的培养基的制备更简单，主要是因为琼脂糖胶凝得更慢并且在生理温度范围内保持液态。此外，将菌落从含有琼脂糖的半固体培养基转移到固体表面组织培养皿比从琼脂转移菌落更有效。 SeaPrep 琼脂糖的储备溶液可以长期保存以供使用。所有这些特征表明，低熔点琼脂糖在制备非贴壁依赖性肿瘤细胞生长的半固体培养基方面比琼脂具有相当大的优势。
The formation of tumor stem cell colonies in vitro has been studied by comparing the growth of three mouse teratocarcinoma derived cell lines and one human teratocarcinoma derived cell line in semi-solid media containing either agar or agarose. We show that agaroses should be used in higher concentrations than agar to obtain comparable results. The maximum number of colonies were obtained in agarose over a broader range of concentrations (1%–4% for SeaPrep and 0.5%–2% for SeaPlaque agarose) than in agar, which allowed anchorage-independent growth of tumor cells only over a narrow concentration range (0.3%–0.5%). Overall, the preparation of media containing agarose was less cumbersome than preparation of agar-containing media, primarily because agaroses gelled more slowly and remained liquid in the physiological temperature range. Furthermore, the transfer of colonies from semi-solid media containing agarose to solid surface tissue culture dishes was much more efficient than the transfer of colonies from agar. The stock solutions of SeaPrep agarose could be kept ready for use for extended periods of time. All these features show that the low melting point agarose has considerable advantages over agar for preparation of semi-solid media for anchorage-independent tumor cell growth.