Stemspan cd34 expansion supplement 10x

Manufactured by STEMCELL

StemSpan™ CD34+ Expansion Supplement (10X) is a cell culture media component designed to support the expansion of CD34+ hematopoietic stem and progenitor cells. It is provided as a concentrated 10X supplement that can be added to appropriate basal media.

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Lab products found in correlation

Stemspan sfem 2, by STEMCELL (3 mentions) Lymphoprep solution, by STEMCELL (1 mentions) Dulbecco s phosphate buffered saline, by Avantor (1 mentions) Cd34 microbead kit ultrapure human, by Miltenyi Biotec (1 mentions) Fix and perm kit, by Thermo Fisher Scientific (1 mentions) Stemspan sfem 2 medium, by STEMCELL (1 mentions) Macs ls column, by Miltenyi Biotec (1 mentions) Fitc annexin 5 apoptosis detection kit 1, by BD (1 mentions) Stemflex media, by Thermo Fisher Scientific (1 mentions) X vivo 15 media, by Lonza (1 mentions) Egm 2 endothelial cell growth medium 2 bulletkit, by Lonza (1 mentions) Pneumacult ex plus medium, by STEMCELL (1 mentions) P3 primary cell nucleofector solution with supplement 1, by Lonza (1 mentions)

Close spelling variants of this product

StemSpan CD34+ expansion supplement (18 citations) CD34+ Expansion Supplement (10 citations) StemSpan (104 citations)

4 protocols using stemspan cd34 expansion supplement 10x

Isolation and Expansion of Human CD34+ Cells

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Citric acid (ACS reagent, ≥99.5%), Diethylentriamine (DETA, 99%), L-Glutamine-Penicillin-Streptomycin solution, Dulbecco’s Phosphate Buffered Saline (DPBS), Float-A-Lyzer dialysis devices (100–500 Da), human serum albumin, EDTA, Selenoprotein-AF647 antibodies and sterile filters (200 nm) were obtained from VWR and antibodies against CD45-FITC/CD34-PE, CD34-APC and the FITC Annexin V Apoptosis Detection Kit I were purchased from BD biosciences. Stem SPAN™ SFEM II medium, Stem SPAN™ CD34⁺ Expansion Supplement (10x) and Lymphoprep™ solution were bought at STEMCELL™ Technologies and microwave reaction vessels were obtained from CEM GmbH. The CD34 MicroBead Kit UltraPure human, MACS LS columns and 30 pre separation filters were purchased from Miltenyi Biotec and the Fix and Perm Kit was bought from Thermo Fisher Scientific. Separation buffer was prepared freshly by supplementing 500 ml DPBS with 1.5 ml 5% human serum albumin and 1.5 ml 50 mM EDTA.

Fasbender S., Zimmermann L., Cadeddu R.P., Luysberg M., Moll B., Janiak C., Heinzel T, & Haas R. (2019). The Low Toxicity of Graphene Quantum Dots is Reflected by Marginal Gene Expression Changes of Primary Human Hematopoietic Stem Cells. Scientific Reports, 9, 12028.

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Cell Culture Media Protocols

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Airway cells: PneumaCult Ex Plus medium (StemCell Technologies #05040) was used for culturing ABCs. Differentiation media was purchased from the University of North Carolina core.
HSPC: StemSpan^™ SFEM II supplemented with StemSpan^™ CD34+ Expansion Supplement (10X) (StemCell Technologies, Catalog # 09720). IPSC: StemFlex^™ media (ThermoFisher Scientific, A3349401). HUVEC: EGM^™−2 Endothelial Cell Growth Medium-2 BulletKit^™ (Lonza, CC-3162). T-cells: X-Vivo 15 media (Lonza; 02-060Q)

Kanke K.L., Rayner R.E., Abel E., Venugopalan A., Suu M., Stack J.T., Nouri R., Guo G., Vetter T.A., Cormet-Boyaka E., Hester M.E, & Vaidyanathan S. (2024). Single-Stranded DNA with Internal Base Modifications Mediates Highly Efficient Gene Insertion in Primary Cells. bioRxiv.

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CD34+ Cell Expansion and Culturing

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G-CSF Mobilized Human Peripheral Blood CD34+ Cells were purchased from StemCell Technologies (Catalog: 70060.1). Cells were thawed by resuspending in 10 mL of RPMI without FBS. The tube was centrifuged at 300 g for 10 minutes and the supernatant removed. Cells were resuspended in StemSpan^™ SFEM II supplemented with StemSpan^™ CD34+ Expansion Supplement (10X) (StemCell Technologies, Catalog # 09720). Cells were plated on 24-well non-treated plates at a density of 100,000 cells/mL and incubated at 37°C for 2–3 days.

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Nucleofection of Hematopoietic Stem Cells

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Hematopoietic stem and progenitor cells were counted after two days and centrifuged at 300 g for 10 minutes. T-cells were counted after 4–6 days in culture. Supernatant was removed and cells were resuspended in P3 Primary Cell Nucleofector^® Solution with Supplement 1 (Lonza, V4XP-3032) at 5–15 million cell/mL. Cell mixture was transferred to Lonza strip at 20 μL per well and electroporated using program DZ100 with solution setting P3. 80 μL of StemSpan^™ SFEM II supplemented with StemSpan^™ CD34+ Expansion Supplement (10X) (StemCell Technologies, Catalog #09720) was added to each well. AAV was added after electroporation, if necessary, at 10⁴ vg/cell. HSPCs were transferred to 24-well non-treated plates at a density of 100,000 cells/mL and incubated at 37°C for 2–3 days. T-cells were cultured at densities of 500,000–1,000,000 cells/mL.

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