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.
Auto macs pro seperator and isolation kit
Manufactured by Miltenyi Biotec
The Auto MACS Pro Separator is a magnetic cell separation system designed for the automated and efficient isolation of specific cell populations from complex samples. The system utilizes magnetic beads coated with antibodies that bind to target cells, allowing for their separation and enrichment. The Auto MACS Pro Separator features a fully automated workflow, streamlining the cell isolation process and ensuring consistent, reproducible results.
Automatically generated - may contain errors
3 protocols using auto macs pro seperator and isolation kit
1
Hematopoietic Stem Cell Isolation and Differentiation
2
Hematopoietic Stem Cell Isolation and Differentiation
Check if the same lab product or an alternative is used in the 5 most similar protocols
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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.
3
Expansion and Differentiation of HSPCs
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