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Epicult basal medium

Manufactured by STEMCELL
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Epicult Basal Medium is a serum-free, xeno-free culture medium designed to support the growth and expansion of human embryonic stem cells and induced pluripotent stem cells. The medium provides the necessary nutrients and growth factors to maintain the undifferentiated state of these cells in culture.

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Lab products found in correlation

Gentle collagenase, by STEMCELL (1 mentions) Zen analysis software, by Zeiss (1 mentions) Matrigel, by STEMCELL (1 mentions) Matrigel, by Corning (1 mentions)

Close spelling variants of this product

EpiCult-B Basal Medium (Human) Epicult-B Basal medium

3 protocols using epicult basal medium

1

Mammary Gland Cell Isolation Protocol

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Mice were injected with the indicated virus in the #3 or 4 mammary glands with no greater than 2 replicates of a single condition per mouse. Individual mammary glands were harvested digested according to Stemcell Technologies gentle collagenase/hyaluronidase protocol. Briefly glands we digested overnight shaking at 37oC in 250 ul gentle collagenase (Stemcell Technologies #07919) in 2.25 ml of complete Basal Epicult media formulated according to manufacture instructions (Epicult Basal Medium Stemcell Technologies #05610, 10% Proliferation Supplement, 5% FBS, 1% Penicillin-Streptomycin, 10 ng/ml EGF, 10 ng/ml bFGF, 0.0004% heparin). Glands were then treated with ammonium chloride and triturated for 2 minutes in pre-warmed trypsin followed by dispase. Cells were stained with CD45, CD31, Ter119, CD49f and EPCAM for luminal and basal cell identification.
Langille E., Al-Zahrani K.N., Ma Z., Liang M., Uuskula-Reimand L., Espin R., Teng K., Malik A., Bergholtz H., El Ghamrasni S., Afiuni-Zadeh S., Tsai R., Alvi S., Elia A., Lü Y., Oh R.H., Kozma K.J., Trcka D., Narimatsu M., Liu J.C., Nguyen T., Barutcu S., Loganathan S.K., Bremner R., Bader G.D., Egan S.E., Cescon D.W., Sørlie T., Wrana J.L., Jackson H.W., Wilson M.D., Witkiewicz A.K., Knudsen E.S., Pujana M.A., Wahl G.M, & Schramek D. (2022). Loss of epigenetic regulation disrupts lineage integrity, induces aberrant alveogenesis and promotes breast cancer. Cancer discovery, 12(12), 2930-2953.
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2

3D Culture of Mammosphere Formation

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Three-dimensional cell culture was performed on 48-well plates coated with Matrigel (356230, Life Sciences, Amsterdam, Netherlands). Wells received 100 µL of Matrigel and plates were left for 30 min at 37 °C for Matrigel gelation. Suspensions of sorted cells were centrifuged at 300 G during 5 min (4 °C) and the cell pellets were resuspended in Epicult medium (Epicult Basal medium, 05602, Stem cell technologies) supplemented with Epicult B supplement (05602, Stem cell technologies), L-Glutamine (2 mM final concentration), hydrocortisone (0.48 µg/mL final concentration) and 2% of Matrigel. Cells were plated at a concentration of 100.000 cells/well and cultured in Epicult medium for 7 days. Cells were observed at 10 × magnification using phase contrast microscopy (Zeiss Axio; Zeiss France, Fougères, France) and images were digitalized. Mammosphere mean diameters were determined from at least 12 frames per conditions using the Zen analysis software (Zeiss, France).
Finot L., Chanat E, & Dessauge F. (2018). Molecular signature of the putative stem/progenitor cells committed to the development of the bovine mammary gland at puberty. Scientific Reports, 8, 16194.
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3

Mammosphere and Matrigel Sphere Assays

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For sphere experiments, MCF10A cells were plated on growth-factor-reduced Matrigel (Corning, Fisher Scientific, Cat#CB-40230C) as described previously (77 (link)) and imaged by bright field after 10 days of sphere growth. Primary mammospheres were isolated from mouse mammary glands and were plated on Corning® Costar® Ultra-Low Attachment 24-Well Plates (CLS3473–24EA) in serum-free Epicult Basal sphere media (Epicult Basal Medium Stemcell Technologies #05610, 10% Proliferation Supplement, 1% Penicillin-Streptomycin, 10 ng/ml EGF, 10 ng/ml bFGF, 0.0004% heparin, + 2% W21 growth supplement). Mammospheres were counted and imaged 10 days after plating.
Langille E., Al-Zahrani K.N., Ma Z., Liang M., Uuskula-Reimand L., Espin R., Teng K., Malik A., Bergholtz H., El Ghamrasni S., Afiuni-Zadeh S., Tsai R., Alvi S., Elia A., Lü Y., Oh R.H., Kozma K.J., Trcka D., Narimatsu M., Liu J.C., Nguyen T., Barutcu S., Loganathan S.K., Bremner R., Bader G.D., Egan S.E., Cescon D.W., Sørlie T., Wrana J.L., Jackson H.W., Wilson M.D., Witkiewicz A.K., Knudsen E.S., Pujana M.A., Wahl G.M, & Schramek D. (2022). Loss of epigenetic regulation disrupts lineage integrity, induces aberrant alveogenesis and promotes breast cancer. Cancer discovery, 12(12), 2930-2953.
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