Evaluating Alpelisib Cytotoxicity on Lymphatic Malformation Cells

Lymphatic malformation-lymphatic endothelial cells (LM-LECs) were maintained as described (Boscolo et al., 2015 (link)) and negative for mycoplasma at the time of these studies. Mycoplasma test was performed using the MycoAlert Mycoplasma Detection Kit (Cat # LT07-218, Lonza) following the manufacturer’s instructions. Real-time analysis of cell viability was performed by using the xCELLigence system RTCA SP (ACEA Biosciences). Briefly, 5 × 10³ LM-LECs per well were seeded in an E-Plate 96 (ACEA Biosciences) and cell proliferation was recorded hourly. When the cells reached the exponential growth phase, new media containing alpelisib at 1, 3, 10, 30, or 100 nM was added and alpelisib cytotoxic effect was recorded hourly. IC₅₀ was calculated by using the dose–response curve function available in the xCELLigence software Version 2.0. Cell index (%) reflects cell viability.

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Shaheen M.F., Tse J.Y., Sokol E.S., Masterson M., Bansal P., Rabinowitz I., Tarleton C.A., Dobroff A.S., Smith T.L., Bocklage T.J., Mannakee B.K., Gutenkunst R.N., Bischoff J., Ness S.A., Riedlinger G.M., Groisberg R., Pasqualini R., Ganesan S, & Arap W. (2022). Genomic landscape of lymphatic malformations: a case series and response to the PI3Kα inhibitor alpelisib in an N-of-1 clinical trial. eLife, 11, e74510.

Publication 2022

MycoAlert Mycoplasma Detection Kit E-Plate 96

Alpelisib Cell Cell proliferation Cell viability Lymphatic endothelial cells Malformation Mycoplasma Rtca

Corresponding Organization :

Other organizations : University of Arizona, Emergency Medicine Foundation, Rutgers Sexual and Reproductive Health and Rights, Rutgers, The State University of New Jersey, Johnson University, New Mexico Cancer Center, University of New Mexico, Markey Cancer Center, University of Kentucky, Boston Children's Hospital, Harvard University

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Variable analysis

independent variables

Alpelisib concentration (1, 3, 10, 30, or 100 nM)

dependent variables

Cell viability (reflected by cell index percentage)

control variables

Lymphatic malformation-lymphatic endothelial cells (LM-LECs) maintained as described in Boscolo et al., 2015
Mycoplasma status (negative for mycoplasma)

controls

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

Annotations

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