Mammosphere Culture and Serial Passage

Mammospheres were cultured under the condition as described previously, specifically, chemically defined cell culture Media, 3dGRO™ Spheroid Medium (S3077, Sigma-Aldrich) was applied, using Ultra-Low attachment plates (CLS3471, Corning) [13 (link), 35 (link)]. After culturing for approximately 7 days, the number of the spheres/2000 cells were counted and scored under an inverted microscope. Sphere formation efficiency = colonies/input cells ×100%. Serial passage was achieved by enzymatic dissociation with Trypsin-EDTA (T4049, Sigma-Aldrich) and Accutase® solution (A6964, Sigma-Aldrich).

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Xiao G., Li X., Li G., Zhang B., Xu C., Qin S., Du N., Wang J., Tang S.C., Zhang J., Ren H., Chen K, & Sun X. (2017). MiR-129 blocks estrogen induction of NOTCH signaling activity in breast cancer stem-like cells. Oncotarget, 8(61), 103261-103273.

Publication 2017

CLS3471 Trypsin-EDTA Accutase® solution

Accutase Cell culture Cells Edta Enzymatic Microscope Trypsin

Corresponding Organization : Huazhong University of Science and Technology

Other organizations : First Affiliated Hospital of Xi'an Jiaotong University, Georgia Regents Medical Center, Augusta University, Tianjin Medical University Cancer Institute and Hospital

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Variable analysis

independent variables

Culturing mammospheres under chemically defined cell culture media, 3dGRO™ Spheroid Medium (S3077, Sigma-Aldrich)
Using Ultra-Low attachment plates (CLS3471, Corning)

dependent variables

Number of spheres per 2000 cells
Sphere formation efficiency (colonies/input cells × 100%)

control variables

Culturing duration (approximately 7 days)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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