Log In Sign up
The largest database of trusted experimental protocols

Bit 9500 medium

Manufactured by STEMCELL
Visit Supplier

BIT 9500 medium is a cell culture medium designed for the maintenance and expansion of human pluripotent stem cells. It provides a defined, serum-free, and animal component-free environment to support the growth and self-renewal of these cells.

Automatically generated - may contain errors

Lab products found in correlation

Iscove modified dulbecco media (imdm), by Corning (3 mentions) Auto macs pro seperator and isolation kit, by Miltenyi Biotec (3 mentions) Auto macs pro separator and isolation kit, by Miltenyi Biotec (2 mentions) Ficoll hypaque, by Miltenyi Biotec (2 mentions) Flt 3 ligand, by STEMCELL (1 mentions) Interleukin 6 (il 6), by STEMCELL (1 mentions) Polybrene, by Merck Group (1 mentions) Thrombopoietin tpo, by Thermo Fisher Scientific (1 mentions) Fms like tyrosine kinase 3 flt3, by Thermo Fisher Scientific (1 mentions) Midi magnetic, by Miltenyi Biotec (1 mentions) Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

BIT 9500 (70 citations) BIT 9500 serum-free medium (2 citations)

6 protocols using bit 9500 medium

1

Isolation and Culture of CD34+ HSPCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
Mononuclear cells were isolated from cord blood using Hetarstach solution (6% Hetastarch in 0.9% NaCl) and Ficoll-Hypaque Plus density centrifugation. CD34+ Hematopoietic Stem and Progenitor Cells (HSPCs) were subsequently purified by positive selection using the Auto MACS Pro Separator and isolation kit (Miltenyi) and were cultured in Iscove’s modified Dulbecco’s medium (IMDM, Cellgro), 20% BIT 9500 medium (Stem Cell Technologies) supplemented with SCF (100 ng ml−1), FLT-3 ligand (10 ng ml−1), IL-6 (20 ng ml−1), and TPO (100 ng ml−1) as the basic culture. All cytokines were purchased from Peprotech, NJ.
Minuesa G., Albanese S.K., Xie W., Kazansky Y., Worroll D., Chow A., Schurer A., Park S.M., Rotsides C.Z., Taggart J., Rizzi A., Naden L.N., Chou T., Gourkanti S., Cappel D., Passarelli M.C., Fairchild L., Adura C., Glickman J.F., Schulman J., Famulare C., Patel M., Eibl J.K., Ross G.M., Bhattacharya S., Tan D.S., Leslie C.S., Beuming T., Patel D.J., Goldgur Y., Chodera J.D, & Kharas M.G. (2019). Small-molecule targeting of MUSASHI RNA-binding activity in acute myeloid leukemia. Nature Communications, 10, 2691.
+ Open protocol
+ Expand
2

Cord Blood CD34+ HSPC Purification and Differentiation

Check if the same lab product or an alternative is used in the 5 most similar protocols
For purification of cord blood CD34+ HSPCs, five to ten mixed CB units (each unit is from a healthy donor) were used. Purified mononuclear cells were obtained from cord blood by using Hespan and Ficoll-Hypaque Plus density centrifugation, followed by positive selection using the Auto MACS Pro Separator and isolation Kit (Miltenyi). CD34+ cells were grown in 80% Iscove’s modified Dulbecco’s medium (IMDM, Cellgro) and 20% BIT 9500 medium (Stem Cell Technologies), supplemented with 100 ng/ml SCF, 10 ng/ml FLT-3 ligand, 20ng/ml IL-6 and 100 ng/ml TPO, which was used as basic media. To differentiate HSPCs into myeloid lineage cells, cells were incubated with myeloid-promoting media that contain 100 ng/ml SCF, 10 ng/ml FLT-3 ligand, 20 ng/ml IL-3, 20 ng/ml IL-6, 20 ng/ml GM-CSF and 20 ng/ml G-CSF. For transduction of CD34+ cells, cells were incubated with high-titer lentiviral supernatant together with 8 μg/ml polybrene (Millipore) and centrifuged at 1600 RPM for 1 hr.
Park S.M., Cho H., Thornton A.M., Barlowe T.S., Chou T., Chhangawala S., Fairchild L., Taggart J., Chow A., Schurer A., Gruet A., Witkin M.D., Kim J.H., Shevach E.M., Krivtsov A., Armstrong S.A., Leslie C, & Kharas M.G. (2018). IKZF2 drives leukemia stem cell self-renewal and inhibits myeloid differentiation. Cell stem cell, 24(1), 153-165.e7.
+ Open protocol
+ Expand
3

Hematopoietic Stem Cell Isolation and Differentiation

Check if the same lab product or an alternative is used in the 5 most similar protocols
All primary patient samples unless otherwise noted were collected under Biospecimen collection/banking study 09-141. The use of the samples for research purposes is covered under biospecimen research protocol 16-354. CD34+ HSPCs were purified from at least 5 or 10 mixed CB units (each unit from one healthy donor) in each purification. Mononuclear cells were first isolated from CB using Hespan and Ficoll-Hypaque Plus density centrifugation, followed by positive selection using the Auto MACS Pro Seperator and isolation Kit (Miltenyi). CD34+ cells were cultured in Iscove’s modified Dulbecco’s medium (IMDM, Cellgro) 20% BIT 9500 medium (Stem Cell Technologies) supplemented with SCF (100 ng/ml), FLT-3 ligand (10 ng/ml), IL-6 (20 ng/ml) and TPO (100 ng/ml) as the basic culture. CD34+ cells were transduced with high-titer retroviral and lentiviral supernatant and 8μg/ml polybrene. To differentiate HSPCs, cells were cultured under the myeloid-promoting conditions: SCF (100 ng/ml), FLT-3 ligands (10 ng/ml), IL-3 (20 ng/ml), IL-6 (20 ng/ml), GM-CSF (20 ng/ml) and G-CSF (20 ng/ml) and the erythroid-promoting conditions: Epo (6 IU/ml) and SCF (100 ng/ml). Cytokines were purchased from Peprotech, NJ.
Vu L.P., Pickering B.F., Cheng Y., Zaccara S., Nguyen D., Minuesa G., Chou T., Chow A., Saletore Y., MacKay M., Schulman J., Famulare C., Patel M., Klimek V.M., Garrett-Bakelman F.E., Melnick A., Carroll M., Mason C.E., Jaffrey S.R, & Kharas M.G. (2017). The N6-methyladenosine (m6A)-forming enzyme METTL3 controls myeloid differentiation of normal and leukemia cells. Nature medicine, 23(11), 1369-1376.
+ Open protocol
+ Expand
4

Isolating and Culturing CD34+ Hematopoietic Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Mononuclear cells were isolated from CB (obtained from the Cleveland Blood Center on a contractual basis) by Ficoll-Hypaque Plus density centrifugation. CD34+ hematopoietic stem and progenitor cells (HSPCs) were purified by positive selection using the Midi-magnetic–activated cell sorting LS columns (130-042-401, Miltenyi Biotec). CD34+ cells were cultured in Iscove modified Dulbecco medium (IMDM) containing 20% BIT 9500 medium (Stem Cell Technologies) supplemented with SCF (100 ng/mL), Fms-like tyrosine kinase 3 (FLT-3; 10 ng/mL), IL-6 (20 ng/mL), and thrombopoietin (TPO; 100 ng/mL; these cytokines were purchased from PeproTech).
Palam L.R., Ramdas B., Pickerell K., Pasupuleti S.K., Kanumuri R., Cesarano A., Szymanski M., Selman B., Dave U.P., Sandusky G., Perna F., Paczesny S, & Kapur R. (2023). Loss of Dnmt3a impairs hematopoietic homeostasis and myeloid cell skewing via the PI3Kinase pathway. JCI Insight, 8(9), e163864.
+ Open protocol
+ Expand
5

Hematopoietic Stem Cell Isolation and Differentiation

Check if the same lab product or an alternative is used in the 5 most similar protocols
All primary patient samples unless otherwise noted were collected under Biospecimen collection/banking study 09-141. The use of the samples for research purposes is covered under biospecimen research protocol 16-354. CD34+ HSPCs were purified from at least 5 or 10 mixed CB units (each unit from one healthy donor) in each purification. Mononuclear cells were first isolated from CB using Hespan and Ficoll-Hypaque Plus density centrifugation, followed by positive selection using the Auto MACS Pro Seperator and isolation Kit (Miltenyi). CD34+ cells were cultured in Iscove’s modified Dulbecco’s medium (IMDM, Cellgro) 20% BIT 9500 medium (Stem Cell Technologies) supplemented with SCF (100 ng/ml), FLT-3 ligand (10 ng/ml), IL-6 (20 ng/ml) and TPO (100 ng/ml) as the basic culture. CD34+ cells were transduced with high-titer retroviral and lentiviral supernatant and 8μg/ml polybrene. To differentiate HSPCs, cells were cultured under the myeloid-promoting conditions: SCF (100 ng/ml), FLT-3 ligands (10 ng/ml), IL-3 (20 ng/ml), IL-6 (20 ng/ml), GM-CSF (20 ng/ml) and G-CSF (20 ng/ml) and the erythroid-promoting conditions: Epo (6 IU/ml) and SCF (100 ng/ml). Cytokines were purchased from Peprotech, NJ.
Vu L.P., Pickering B.F., Cheng Y., Zaccara S., Nguyen D., Minuesa G., Chou T., Chow A., Saletore Y., MacKay M., Schulman J., Famulare C., Patel M., Klimek V.M., Garrett-Bakelman F.E., Melnick A., Carroll M., Mason C.E., Jaffrey S.R, & Kharas M.G. (2017). The N6-methyladenosine (m6A)-forming enzyme METTL3 controls myeloid differentiation of normal and leukemia cells. Nature medicine, 23(11), 1369-1376.
+ Open protocol
+ Expand
6

Expansion and Differentiation of HSPCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
CD34+ HSPCs were purified from at least 10 mixed CB units (each unit from one healthy donor) in each purification. After Hespan and Ficoll-Hypaque Plus density centrifugation, mononuclear cells from CB units were performed positive selection using the Auto MACS Pro Seperator and isolation Kit (Miltenyi). CD34+ cells were cultured in basic medium containing Iscove’s modified Dulbecco’s medium (IMDM, Cellgro) 20% BIT 9500 medium (Stem Cell Technologies) supplemented with SCF (100 ng/ml), FLT-3 ligand (10 ng/ml), IL-6 (20 ng/ml) and TPO (100 ng/ml). To differentiate HSPCs, cells were cultured under the myeloid-promoting conditions: SCF (100 ng/ml), FLT-3 ligands (10 ng/ml), IL-3 (20 ng/ml), IL-6 (20 ng/ml), GM-CSF (20 ng/ml) and G-CSF (20 ng/ml) or the erythroid-promoting conditions: Epo (6 IU/ml) and SCF (100 ng/ml). Cytokines were purchased from Peprotech, NJ.
Cheng Y., Xie W., Pickering B.F., Chu K.L., Savino A.M., Yang X., Luo H., Nguyen D.T., Mo S., Barin E., Velleca A., Rohwetter T., Patel D.J., Jaffrey S.R, & Kharas M.G. (2021). N6-methyladenosine on mRNA facilitates a phase-separated nuclear body that suppresses myeloid leukemic differentiation. Cancer cell, 39(7), 958-972.e8.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!