Expansion and Differentiation of HUDEP-2 Cells

HUDEP-2 cells were used as previously described^{3 (link),31 (link)}, which tested negative for mycoplasma contamination. HUDEP-2 cells were expanded in SFEM (Stem Cell Technologies) supplemented with 100 ng/mL stem cell factor (R&D), 3 UI/mL erythropoietin (Amgen), 10⁻⁶ M dexamethasone (Sigma), 1 µ/mL of doxycycline (Sigma), and 2% penicillin-streptomycin (Thermo Fisher). HUDEP-2 cells were differentiated in Iscove’s Modified Dulbecco’s Medium (IMDM) supplemented with 330 µg/mL holo-human transferrin (Sigma), 10 µg/mL recombinant human insulin (Sigma), 2 IU/mL heparin (Sigma), 5% human solvent detergent pooled plasma AB (Rhode Island Blood Center), 3 IU/mL erythropoietin (Amgen), 100 ng/mL human stem cell factor (SCF) (R&D), 1 µg/mL doxycycline (Sigma), 1% L-glutamine (Life Technologies), and 2% penicillin/streptomycin (Life Technologies).

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Canver M.C., Lessard S., Pinello L., Wu Y., Ilboudo Y., Stern E.N., Needleman A.J., Galactéros F., Brugnara C., Kutlar A., McKenzie C., Reid M., Chen D.D., Das P.P., Cole M., Zeng J., Kurita R., Nakamura Y., Yuan G.C., Lettre G., Bauer D.E, & Orkin S.H. (2017). Variant-aware saturating mutagenesis using multiple Cas9 nucleases identifies regulatory elements at trait-associated loci. Nature genetics, 49(4), 625-634.

Publication 2017

stem cell factor erythropoietin dexamethasone doxycycline penicillin-streptomycin holo-human transferrin recombinant human insulin heparin human stem cell factor (SCF) L-glutamine

Blood Cells Detergent Dexamethasone Doxycycline Erythropoietin Glutamine Heparin Human Human stem cell factor Insulin Mycoplasma Penicillin Plasma Solvent Stem cell Stem cell factor Streptomycin Transferrin

Corresponding Organization :

Other organizations : Dana-Farber Cancer Institute, Boston Children's Hospital, Harvard Stem Cell Institute, Harvard University, Montreal Heart Institute, Université de Montréal, Massachusetts General Hospital, Institut Mondor de Recherche Biomédicale, Assistance Publique – Hôpitaux de Paris, Augusta University Health, University of the West Indies, Japanese Red Cross Society, Japan, RIKEN BioResource Research Center

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Variable analysis

independent variables

HUDEP-2 cell expansion media composition
HUDEP-2 cell differentiation media composition

dependent variables

HUDEP-2 cell growth and differentiation

control variables

HUDEP-2 cells tested negative for mycoplasma contamination

controls

Positive control: HUDEP-2 cells expanded and differentiated according to previously described protocols (references 3 and 31)
Negative control: Not explicitly mentioned

Annotations

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