Lipid tethering of breast tumor cells reduces cell aggregation during mammosphere formation.
Lipid tethering of breast tumor cells reduces cell aggregation during mammosphere formation.
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乳腺肿瘤细胞的脂质栓系减少了乳腺球形成过程中的细胞聚集。
DOI：
10.1038/s41598-021-81919-9
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发表时间：
2021-02-05
影响因子：
4.6
通讯作者：
Martin SS
中科院分区：
文献类型：
--
作者：
Bhandary L;Bailey PC;Chang KT;Underwood KF;Lee CJ;Whipple RA;Jewell CM;Ory E;Thompson KN;Ju JA;Mathias TM;Pratt SJP;Vitolo MI;Martin SS
乳房试验在体外被广泛用于识别潜在的致癌干细胞，这些干细胞可以在自由漂浮的条件下克隆繁殖形成球体。然而，传统的乳房小球试验不可避免地会引入细胞聚集，这会干扰真正的乳房小球形成效率的测量。我们开发了一种方法来减少肿瘤细胞聚集，并增加观察到的乳房形成的起源是克隆的可能性。将单个肿瘤细胞系在脂质锚上可以防止细胞漂移，同时保持自由漂移的特性。这使得能够实时监测单个肿瘤细胞分裂形成乳房的过程。监测系留的乳腺癌细胞提供了详细的大小信息，这些信息与之前公布的单细胞跟踪数据直接相关。我们观察到，与传统的低附着性钢板中只有37%的球相比，脂质涂层孔中7day球的71%的μm在50到150um之间。当MCF7-GFP和MCF7-mCherry细胞等比例混合接种时，在脂膜覆盖孔中65%的乳房细胞显示单色表达，而在低附着孔中只有32%显示单色表达。这些结果表明，使用脂质拴系用于乳房生长测定，可以减少细胞聚集的混杂因素，增加克隆乳房的形成。
Mammosphere assays are widely used in vitro to identify prospective cancer-initiating stem cells that can propagate clonally to form spheres in free-floating conditions. However, the traditional mammosphere assay inevitably introduces cell aggregation that interferes with the measurement of true mammosphere forming efficiency. We developed a method to reduce tumor cell aggregation and increase the probability that the observed mammospheres formed are clonal in origin. Tethering individual tumor cells to lipid anchors prevents cell drift while maintaining free-floating characteristics. This enables real-time monitoring of single tumor cells as they divide to form mammospheres. Monitoring tethered breast cancer cells provided detailed size information that correlates directly to previously published single cell tracking data. We observed that 71% of the Day 7 spheres in lipid-coated wells were between 50 and 150 μm compared to only 37% in traditional low attachment plates. When an equal mixture of MCF7-GFP and MCF7-mCherry cells were seeded, 65% of the mammospheres in lipid-coated wells demonstrated single color expression whereas only 32% were single-colored in low attachment wells. These results indicate that using lipid tethering for mammosphere growth assays can reduce the confounding factor of cell aggregation and increase the formation of clonal mammospheres.
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影响因子：
10.5
作者：
Dontu, G;Abdallah, WM;Wicha, MS
通讯作者：
Wicha, MS
影响因子：
11.2
作者：
Huang EH;Hynes MJ;Zhang T;Ginestier C;Dontu G;Appelman H;Fields JZ;Wicha MS;Boman BM
通讯作者：
Boman BM
影响因子：
5.8
作者：
Bailey PC;Lee RM;Vitolo MI;Pratt SJP;Ory E;Chakrabarti K;Lee CJ;Thompson KN;Martin SS
通讯作者：
Martin SS
影响因子：
1.2
作者：
Lombardo, Ylenia;de Giorgio, Alexander;Castellano, Leandro
通讯作者：
Castellano, Leandro
影响因子：
11.2
作者：
Boggs AE;Vitolo MI;Whipple RA;Charpentier MS;Goloubeva OG;Ioffe OB;Tuttle KC;Slovic J;Lu Y;Mills GB;Martin SS
通讯作者：
Martin SS