Stemspan erythroid expansion supplement

Manufactured by STEMCELL

Sourced in Canada

StemSpan Erythroid Expansion Supplement is a cell culture medium component designed to support the expansion of erythroid cells in vitro. It provides the necessary growth factors and nutrients to promote the proliferation and differentiation of erythroid progenitor cells.

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Lab products found in correlation

Stemspan sfem 2 medium, by STEMCELL (4 mentions) Cytotune sendai viral vector kit, by Thermo Fisher Scientific (1 mentions) Stemspan sfem 2, by STEMCELL (1 mentions) Rosettesep human progenitor cell basic pre enrichment antibody cocktail, by STEMCELL (1 mentions)

Close spelling variants of this product

StemSpan (104 citations) Erythroid Expansion Supplement (10 citations) StemSpan-SFEM II with 100X StemSpan Erythroid Expansion Supplement (2 citations)

6 protocols using stemspan erythroid expansion supplement

Erythroid Differentiation of hPSC-derived Progenitors

Cited 1 time

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Erythroid differentiation was performed using the StemSpan Erythroid Expansion Supplement (STEMCELL Technologies, 02692) following the manufacturer’s recommendations. In brief, day 10 hPSC-derived HLF+ HOXA+ hematopoietic progenitors were collected, counted, and seeded in StemSpan SFEM II (STEMCELL Technologies, 09655) supplemented with the StemSpan Erythroid Expansion Supplement at a density of 1x10⁴-1x10⁵ cells/mL. On day 3, cells were supplemented with an equal volume of complete media. On day 7 and 10, cells were harvested and replated in complete media at a density of 1x10⁵ cells/mL. On day 14, cells were harvested and processed for flow cytometry.
High-performance liquid chromatography (HPLC) to detect hemoglobin tetramers was performed as previously described.²⁰⁶ (link)

Fowler J.L., Zheng S.L., Nguyen A., Chen A., Xiong X., Chai T., Chen J.Y., Karigane D., Banuelos A.M., Niizuma K., Kayamori K., Nishimura T., Cromer M.K., Gonzalez-Perez D., Mason C., Liu D.D., Yilmaz L., Miquerol L., Porteus M.H., Luca V.C., Majeti R., Nakauchi H., Red-Horse K., Weissman I.L., Ang L.T, & Loh K.M. (2024). Lineage-tracing hematopoietic stem cell origins in vivo to efficiently make human HLF+ HOXA+ hematopoietic progenitors from pluripotent stem cells. Developmental Cell, 59(9), 1110-1131.e22.

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Erythroid Expansion Modulation by βIP

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To demonstrate whether βIP had an effect on erythropoiesis, we analyzed erythroid expansion with and without the novel βIP (1 mM). Peripheral blood mononuclear cells (PBMCs) from a healthy adult donor were purchased from StemCell Technologies (Vancouver, Canada). PBMC were seeded at a density of 0.5 × 10⁶ cells/ml in StemSpan SFEM II medium (StemCell Technologies) supplemented with or without StemSpan Erythroid Expansion Supplement (StemCell Technologies) and 1% (vol/vol) P/S. Cells were cultured for 8 days at 37°C in 5% CO2 incubator with the media being changed every two days [29 (link)]. PBMCs are treated with peptide inhibitor (25μM) from Day 2.

Kilar C.R., Sekharan S., Sautina L., Diao Y., Keinan S., Shen Y., Bungert J., Mohandas R, & Segal M.S. (2018). Computational Design and Experimental Characterization of a Novel β-Common Receptor Inhibitory Peptide. Peptides, 104, 1-6.

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Generation of PTPN11 Q510P iPSCs

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Patient PBMCs (LS PTPN11 Q510P, #9915) were obtained from Dr. Amy Roberts at Boston Children’s Hospital. One million patient PBMCs were cultured in one well of 12-well tissue culture plate with 1ml StemSpan™ SFEM II medium supplemented with StemSpan™ Erythroid Expansion Supplement (StemCell Technologies) for expansion of human erythroid cells for 10 days and then reprogrammed into iPSCs using the non-integrating CytoTune–Sendai viral vector kit (A16517, Thermo Fisher Scientific) following the method described previously [4 (link), 5 (link)]. On day 4, cells were re-plated onto a Matrigel-coated dish in E8-media based reprogramming media, and fed every other day until day 20 when individual colonies were passaged by the EDTA dissociation method into separate wells in E8 medium. The selected iPSC colonies were further cultured for more than 15 passages before downstream application.

Li R., Baskfield A., Lin Y., Beers J., Zou J., Liu C., Jaffré F., Roberts A.E., Ottinger E.A., Kontaridis M.I, & Zheng W. (2018). Generation of an induced pluripotent stem cell line (TRNDi003-A) from a Noonan syndrome with multiple lentigines (NSML) patient carrying a p.Q510P mutation in the PTPN11 gene. Stem cell research, 34, 101374.

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Isolation and Expansion of Patient-Derived iPSCs

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To isolate mononuclear cells, 10 ml of peripheral blood was collected from a proband patient with a PDE6B mutation and an unaffected volunteer with no retinal disease and no mutation at any RP related genes following the instructions for Lymphoprep (Cat. #07851; Stemcell Technologies, Norway). Cells were maintained in StemSpan SFEM II medium (Cat. #09605; Stemcell Technologies, Canada) supplied with StemSpan Erythroid Expansion Supplement (Cat. #02692; Stemcell Technologies, Canada) for seven days for cell expansion. Both the patient and control volunteers were signed informed content, which has been approved by the Ethics Committee of the Eye Hospital of Wenzhou Medical University. The patient and control iPSCs were included in the hiPSC bank of Institute of Stem Cell Research, Wenzhou Medical University and designated as 502-PBMC-PDE6B-01 and 502-PBMC-HEALTHY-01.

Gao M.L., Lei X.L., Han F., He K.W., Jin S.Q., Zhang Y.Y, & Jin Z.B. (2020). Patient-Specific Retinal Organoids Recapitulate Disease Features of Late-Onset Retinitis Pigmentosa. Frontiers in Cell and Developmental Biology, 8, 128.

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Isolation and Expansion of Erythroid Progenitors

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Peripheral blood was collected from the donor into EDTA-containing blood collection tube and treated with RosetteSep™ Human Progenitor Cell Basic Pre-Enrichment antibody cocktail according to the manufacturer’s instructions (StemCell Technologies Catalog#15226). After PBMCs separation and isolation, 1 million cells were cultured for 8 days in StemSpan™ SFEM II medium (StemCell Technologies Catalog #09605) supplemented with 1X StemSpan™ Erythroid Expansion Supplement (StemCell Technologies Catalog #02692).

Alowaysi M., Lehmann R., Al-Shehri M., Baadhaim M., Alzahrani H., Aboalola D., Zia A., Malibari D., Daghestani M., Alghamdi K., Haneef A., Jawdat D., Hakami F., Gomez-Cabrero D., Tegner J, & Alsayegh K. (2023). HLA-based banking of induced pluripotent stem cells in Saudi Arabia. Stem Cell Research & Therapy, 14, 374.

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Erythroid Differentiation of Cryopreserved CD34+ Cells

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Cryopreserved human bone marrow CD34⁺ cells were purchased from STEMCELL Technologies (Catalog No. 70002.3; Vancouver, Canada). Frozen bone marrow CD34⁺ cells were thawed, washed once with StemSpan™ SFEM II medium (Catalog No. 09605; STEMCELL Technologies), suspended in StemSpan™ SFEM II medium containing StemSpan™ Erythroid Expansion Supplement (Catalog No. 02692; STEMCELL Technologies), and cultured for 7 days. CD34⁺ cells were then harvested, washed once with Dulbecco's (D)‐PBS, and suspended at 0.5 x 10⁶ cells/mL in human whole blood to be used in the EPO stimulation assay.
Human whole blood was collected from 13 healthy donors (seven males and six females) via venipuncture into Vacutainer collection tubes containing sodium heparin, in accordance with Pfizer protocols (Protocol No. GOHW RDP‐01) approved by the Shulman Institutional Review Board. Blood was warmed to 37°C prior to use.

Dowty M.E., Lin T.H., Jesson M.I., Hegen M., Martin D.A., Katkade V., Menon S, & Telliez J. (2019). Janus kinase inhibitors for the treatment of rheumatoid arthritis demonstrate similar profiles of in vitro cytokine receptor inhibition. Pharmacology Research & Perspectives, 7(6), e00537.

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