Cryopreserved primary CD34+ cells were recovered from liquid nitrogen as previously described10 (link) and cultured overnight in a humidified incubator at 37 °C with 5% CO2 in SFM consisting of Iscove Modified Dulbecco Medium (Sigma-Aldrich) containing serum substitute (bovine serum albumin, insulin, transferrin [BIT]; Stem Cell Technologies), 0.1 μM 2-mercaptoethanol (Sigma-Aldrich), penicillin-streptomycin, L-glutamine plus a high concentration growth factor (5GF) cocktail containing 100 ng/mL Flt3-ligand (Flt3-L), 100 ng/mL stem cell factor (SCF), 20 ng/mL interleukin-3 (IL-3), 20 ng/mL IL-6 (Stem Cell Technologies) and 20 ng/mL granulocyte-colony stimulating factor (G-CSF; Chugai Pharma) to maximize cell recovery. In selected experiments following overnight culture, cells were washed and cultured in SFM supplemented with 5GF cocktail, a physiological growth factor cocktail comprising 5 ng/mL Flt3-L, 5 ng/mL SCF, 1 ng/mL IL-3, 1 ng/mL IL-6 and 1 ng/mL G-CSF (LGF) or SFM alone.
Iscove modified dulbecco medium
Manufactured by Merck Group
Iscove Modified Dulbecco Medium (IMDM) is a cell culture medium formulation developed for the growth and maintenance of various cell types, including hematopoietic and lymphoid cells. It is designed to provide a balanced set of nutrients and growth factors necessary for supporting the in vitro cultivation of these cell types.
Automatically generated - may contain errors
Lab products found in correlation
Bovine serum albumin (bsa), by STEMCELL (1 mentions)
2 mercaptoethanol, by Merck Group (1 mentions)
Interleukin 6 (il 6), by STEMCELL (1 mentions)
Panc 1, by BNCC (1 mentions)
Hs766t, by BNCC (1 mentions)
Sw1990, by BNCC (1 mentions)
Cfpac 1, by BNCC (1 mentions)
Cfpac 1, by Merck Group (1 mentions)
Sw1990, by Merck Group (1 mentions)
Leibovitz l 15 medium, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
M3 base f culture medium, by INCELL (1 mentions)
2 protocols using iscove modified dulbecco medium
1
Cryopreserved CD34+ Cell Culture Optimization
2
Culturing Human Pancreatic Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Human pancreatic ductal cell line (HPNE) and PC cell lines, including PANC-1, SW1990,
HS766T, and CFPAC-1, were purchased from BeNa Culture Collection (Beijing, China) and
cultured in incubator with 100% humidity and 5% CO2 at 37°C. The HPNE cells
were grown in Dulbecco modified Eagle medium (DMEM; Sigma, St Louis, Missouri) containing
1 volume of M3 Base F culture medium (InCell Corp, San Antonio, Texas), 3 volumes of
glucose-free DMEM, 10% fetal bovine serum (FBS; (Invitrogen, Carlsbad, California), 5.5 mM
glucose, 10 ng/mL epidermal growth factor (EGF), and 50 µg/mL gentamycin. The DMEM medium
with 10% FBS was applied to culture PANC-1 and HS766T cells. SW1990 cells were maintained
in Leibovitz L-15 Medium (Sigma). Furthermore, CFPAC-1 cells were cultured in Iscove
Modified Dulbecco Medium (Sigma) adding 10% FBS.
HS766T, and CFPAC-1, were purchased from BeNa Culture Collection (Beijing, China) and
cultured in incubator with 100% humidity and 5% CO2 at 37°C. The HPNE cells
were grown in Dulbecco modified Eagle medium (DMEM; Sigma, St Louis, Missouri) containing
1 volume of M3 Base F culture medium (InCell Corp, San Antonio, Texas), 3 volumes of
glucose-free DMEM, 10% fetal bovine serum (FBS; (Invitrogen, Carlsbad, California), 5.5 mM
glucose, 10 ng/mL epidermal growth factor (EGF), and 50 µg/mL gentamycin. The DMEM medium
with 10% FBS was applied to culture PANC-1 and HS766T cells. SW1990 cells were maintained
in Leibovitz L-15 Medium (Sigma). Furthermore, CFPAC-1 cells were cultured in Iscove
Modified Dulbecco Medium (Sigma) adding 10% FBS.
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