Stempro 34

Manufactured by Thermo Fisher Scientific

Sourced in United States

StemPro-34 is a serum-free medium designed for the culture and expansion of human hematopoietic stem and progenitor cells. It contains a proprietary blend of nutrients, growth factors, and other components to support the growth and differentiation of these cell types.

Automatically generated - may contain errors

Lab products found in correlation

74 protocols using stempro 34

Isolation and Differentiation of Hematopoietic Cells

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

T cells, B cells, Mph and DCs were sorted by flow cytometry using markers TCRβ, CD4, CD8, CD11c, CD11b, CD19, CD45R, MHC II as described before (12 (link)). Purity of sorted cells was generally >98% and viability was >97% as determined by 7-AAD staining.
Erythrocytes, normoblasts, reticulocytes and red blood cells were generated using a mouse adapted protocol for the long-term ex vivo erythroid differentiation culture protocol described by Giarratana et al (17 (link)) and Konstantinidis et al (18 (link)). Briefly, low-density bone marrow cells were cultured in erythroblast growth medium (StemPro-34 with 2.6% StemPro-34 supplement; Invitrogen), 20% BIT 9500 (StemCell Technologies), 900 ng/mL ferrous sulfate, 90 ng/mL ferrous nitrate, 10⁻⁶M hydrocortisone, penicillin/streptomycin, L-glutamine), in 3 subsequent phases. For the proliferative phase (days 1–5) cells were expanded with 100 ng/mL SCF, 5 ng/mL IL-3, and 2 IU/mL human erythropoietin (Amgen). In the differentiation step (days 6–7), the cells were supplemented with only erythropoietin in fibronectin coated plates. For enucleation (day 8–9) cells were grown without cytokines. Cells were sorted based on size gating and nucleotide staining using different combinations of Syto-16, Draq5, Mitotracker Red and MitotrackerGreen (Invitrogen).

Klarquist J., Hennies C.M., Lehn M.A., Reboulet R.A., Feau S, & Janssen E.M. (2014). STING-mediated DNA sensing promotes antitumor and autoimmune responses to dying cells. Journal of immunology (Baltimore, Md. : 1950), 193(12), 6124-6134.

+ Open protocol

+ Expand

Directed Differentiation of hPSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

To observe the effects of the two (2D and 3D) culture systems on differentiation, neither Wnt pathway activators nor inhibitors were added to the media as conventionally done (See Table 1). The differentiation of the hPSCs took place in 12-well plates (Nunc) treated with MG for the 2D and in ultra-low attachment non-treated plates for 3D. Mesodermal differentiation proceeded in Stempro 34 (Sigma-Aldrich) medium supplemented with 10× ITS (Gibco), 40 ng/mL BMP4, 10ng/mL FGF2 (Preprotech), 50 μg/mL Ascorbic Acid (Sigma-Aldrich), 100× Glutamax (Gibco) and Gentamicin (Gibco) from day 0 to day 4. On day 4, the medium was changed to Stempro 34 with 10× ITS, 100× Glutamax, Gentamicin, 10 ng/mL FGF, 10 ng/mL VEGF; 10 ng/mL of IL6 and 2 U/mL EPO (Preprotech) for three more days (days 4 to 6). On day 6 the medium was replaced with hematopoietic differentiation Stempro 34 medium containing the above supplements plus 50 ng/mL SCF, 5 ng/mL IL7, 5 ng/mL Flt3 and 10 ng/mL IL3 (Preprotech Rocky Hill, USA). CHIR99021 (3 μM) and IWR-1(1 µM) (Sigma), were added only when indicated.

Mora-Roldan G.A., Ramirez-Ramirez D., Pelayo R, & Gazarian K. (2021). Assessment of the Hematopoietic Differentiation Potential of Human Pluripotent Stem Cells in 2D and 3D Culture Systems. Cells, 10(11), 2858.

+ Open protocol

+ Expand

Directed Differentiation of iPSCs into ECs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Regarding endothelial differentiation, on day 0, iPSC colonies were dissociated into single cells with Accutase (Sigma). Cells were resuspended in basic medium containing StemPro34 (Invitrogen), L-glutamine (Invitrogen), transferrin (Roche), monothioglycerol (Sigma), and ascorbic acid (Sigma), and Y-27632 (naclai tesque), BMP-4 (Peprotech), and Matrigel (BD) were added to form EBs. On day 1, the medium was supplemented with Activin A (R&D Systems) and BMP4. On day 4, EBs were seeded in Matrigel-coated dishes, and the medium was supplemented with VEGF (R&D Systems) for EC expansion. On day 14, the EBs were harvested using Accutase (Sigma-Aldrich), and CD31 positive cells were positively sorted using magnetic-activated cell sorting using anti-CD31 (Miltenyi) (Fig. 7a).

Sekine S.I., Kaneko M., Tanaka M., Ninomiya Y., Kurita H., Inden M., Yamada M., Hayashi Y., Inuzuka T., Mitsui J., Ishiura H., Iwata A., Fujigasaki H., Tamaki H., Tamaki R., Kito S., Taguchi Y., Tanaka K., Atsuta N., Sobue G., Kondo T., Inoue H., Tsuji S, & Hozumi I. (2019). Functional evaluation of PDGFB-variants in idiopathic basal ganglia calcification, using patient-derived iPS cells. Scientific Reports, 9, 5698.

+ Open protocol

+ Expand

Monocyte Differentiation into Fibrocytes

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Low-density BM cells were cultured in conditions that promote differentiation of monocytes to fibrocytes (Pilling et al., 2003 (link)) as described previously (Pilling et al., 2009 (link)). In brief, CD14⁺ monocytes purified from low-density BM cells (purity >97%) were cultured with serum-free medium (StemPro-34; Invitrogen) supplemented with 10 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (Sigma-Aldrich), 1× nonessential amino acids (Sigma-Aldrich), 1 mM sodium pyruvate (Sigma-Aldrich), 2 mM glutamine (Invitrogen), 100 U/ml penicillin, 100 µg/ml streptomycin, and 1× ITS-3 (containing 10 µg/ml insulin, 5 µg/ml transferrin, 5 ng/ml sodium selenite, 0.5 mg/ml albumin, 5 µg/ml oleic acid, and 5 µg/ml linoleic acid [Sigma-Aldrich]), either in flat-bottomed 96-well plates or in Lab-Tek microscope chamber slides (CC2 slides; Thermo Fisher Scientific). Culture plates and slides were maintained at 37°C in humidified air supplemented with 5% CO₂.
Cells were visualized using a phase-contrast microscope (ELWD 0.3; 10/0.25 objective lens; Nikon), photographed using a digital camera (D40; Nikon), and counted as previously described (Pilling et al., 2003 (link); 2009). The number of fibrocytes generated in culture varied by patient sample but ranged from 200 to 400 cells.

Verstovsek S., Manshouri T., Pilling D., Bueso-Ramos C.E., Newberry K.J., Prijic S., Knez L., Bozinovic K., Harris D.M., Spaeth E.L., Post S.M., Multani A.S., Rampal R.K., Ahn J., Levine R.L., Creighton C.J., Kantarjian H.M, & Estrov Z. (2016). Role of neoplastic monocyte-derived fibrocytes in primary myelofibrosis. The Journal of Experimental Medicine, 213(9), 1723-1740.

+ Open protocol

+ Expand

Embryoid Body Generation from hESCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

To generate EBs, we treated hESCs with collagenase B (1 mg/mL; Roche) for 20 min. Cells were gently scraped with a cell scraper to form small aggregates (10–20 cells). Aggregates were resuspended in StemPro-34 (Invitrogen), supplemented with L-glutamine (2 mM), ascorbic acid (1 mM), monothioglycerol (4 × 10⁻⁴ M; Sigma-Aldrich), and transferrin (150 mg/mL; Roche). BMP4 (10 ng/mL; R&D), basic fibroblast growth factor (5 ng/mL; Peprotech), SB431542 (6 μM; Tocris), vascular endothelial growth factor (15 ng/mL; R&D), interleukin-6 (IL-6) (10 ng/mL; R&D), insulin-like growth factor 1 (25 ng/mL; R&D), IL-11 (5 ng/mL; R&D), stem cell factor (SCF) (50 ng/mL; Miltenyi), erythropoietin (2 U/mL), thrombopoietin (30 ng/mL; R&D), IL-3 (30 ng/mL; R&D), and FMS-like tyrosine kinase 3 ligand (FLT3L) (10 ng/mL; Miltenyi) were added as indicated (see Figure 3A). Cultures were maintained in a 5% CO₂/5% O₂/90% N₂ environment. On the day of assay, EBs were harvested and dissociated to single cells by a 40-min treatment with 0.2% collagenase IV. Afterward, 1 mL of medium with serum was added and the EBs were dissociated to single cells by passaging six times through a 20-gauge needle.

Li Y., Brauer P.M., Singh J., Xhiku S., Yoganathan K., Zúñiga-Pflücker J.C, & Anderson M.K. (2017). Targeted Disruption of TCF12 Reveals HEB as Essential in Human Mesodermal Specification and Hematopoiesis. Stem Cell Reports, 9(3), 779-795.

+ Open protocol

+ Expand

Differentiation of hESCs into Endothelial Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Prior to formation of EBs, the hESCs were maintained on matrigel for at least 1 generation. To form EBs, hESC colonies were dissociated with dispase (1 mg/mL; Invitrogen) and gently scraped off from the wells. The cell clusters were pipetted to about 20 to 100 cells in size and then cultured in mTeSR1 with 10 μmol/L Y27632 (Selleck Chemicals, Houston, TX). At the second day, the medium was changed to StemPro‐34 (Invitrogen) supplemented with 1 mmol/L ascorbic acid, 2 mmol/L l‐glutamine, and transferrin (Sigma‐Aldrich). Activin A, BMP4 (BD Biosciences), and bFGF were added on this day. After 3 days of culture, human‐VEGF and XAV939 were added for another 4 days. Then the supplemental factors were changed to VEGF (Pepro Tech, Rocky Hill, NJ) and bFGF, and cells were harvested at the 15th or 30th day.

Qiu X., Liu Y., Zhang Y., Guan Y., Jia Q., Wang C., Liang H., Li Y., Yang H., Qin Y., Huang S., Zhao X, & Jing Q. (2017). Rapamycin and CHIR99021 Coordinate Robust Cardiomyocyte Differentiation From Human Pluripotent Stem Cells Via Reducing p53‐Dependent Apoptosis. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 6(10), e005295.

+ Open protocol

+ Expand

Isolation and Culture of CD34+ Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Following informed consent umbilical cord blood (CB) was collected from healthy newborns and was used within 4 h after delivery. Mononuclear blood cells were separated by centrifugation on a layer of Ficoll-Paque Leucosep (Greiner Bio-One GmbH, Austria). Human CD34+ stem cells were isolated using the CD34 microbead isolation kit from Miltenyi Biotec (Germany) according to the manufacturer’s instructions. The CD34+ cells were cultured in serum-free medium (Stempro34, Invitrogen) containing interleukin-3 (IL-3) and stem cell factor (SCF) at 1 ng/mL and thrombopoietin (TPO) at 50 ng/mL, with medium changes twice weekly. After the first 6 days of culture, the growth medium was supplemented with only TPO at 50 ng/mL. The study was approved by the Ethics Committee of the Medical University of Vienna, Austria.

Ramanathan G, & Mannhalter C. (2016). Increased expression of transient receptor potential canonical 6 (TRPC6) in differentiating human megakaryocytes. Cell biology international, 40(2), 223-231.

+ Open protocol

+ Expand

Directed Differentiation of iPSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

iPSC colonies were detached using colagenase type IV (Gibco) for 30 min at 37°C, washed and centrifuged at 200× g, resuspended in differentiation media composed by KO-DMEM (Gibco) supplemented with 20% non-heat-inactivated FBS (Biowhitaker), 1% NEAA (Lonza; Biowhitaker), L-Glu (1 mM; Invitrogen), β-mercaptoethanol (0.1 mM; Gibco) and hrBMP4 (0.5 ng/ml; Prepotech) and plated in ultra-low attachment plates (Costar). After 2 days, media were replaced by Stempro 34 (Invitrogen) supplemented with 0.5% pen/streptomicin, L-Glu (2 mM; Invitrogen), MTG (40 mM; Sigma), ascorbic acid (50 μg/ml; Invitrogen), hrSCF, hrFlt3 ligand and TPO (100 ng/ml; EuroBioSciences), hrIL3 (10 ng/ml; Biosource), hrIL6 (10 ng/ml; Prepotech), hrBMP4 (50 ng/ml; Prepotech), Wnt11 (200 ng/ml; R&D), and rhVEGF (5 ng/ml; Prepotech). Media were changed every 3–4 days. At day 7, media were replaced by fresh media where rhWnt-11 was substituted by rhWnt-3a (200 ng/ml; R&D). Media were changed every 3–4 days. At day 14 and 21, immunophenotypic analysis of the differentiated cells was performed by flow cytometry, and colony-forming unit assays were conducted (See Supplementary Methods).

Rio P., Baños R., Lombardo A., Quintana-Bustamante O., Alvarez L., Garate Z., Genovese P., Almarza E., Valeri A., Díez B., Navarro S., Torres Y., Trujillo J.P., Murillas R., Segovia J.C., Samper E., Surralles J., Gregory P.D., Holmes M.C., Naldini L, & Bueren J.A. (2014). Targeted gene therapy and cell reprogramming in Fanconi anemia. EMBO Molecular Medicine, 6(6), 835-848.

+ Open protocol

+ Expand

Generating iCAR-NK/ILC from T-iPSC

Check if the same lab product or an alternative is used in the 5 most similar protocols

G2‐CAR‐transduced QHJI01s04 was differentiated into a hematopoietic precursor through the feeder‐free embryoid body formation method. Undifferentiated T‐iPSC colonies were treated with TrypLE select (Gibco) for 8 minutes and transferred to low‐attachment plates and incubated overnight in Stemfit AK03N containing 10 μmol/L Y‐27632 to allow for the formation of embryoid bodies (EB). The EB were collected and transferred to EB medium (StemPro‐34 [Invitrogen], 2 mmol/L l‐glutamine, 400 μmol/L monothioglycerol, 50 μg/mL ascorbic acid‐2‐ phosphate and insulin‐transferrin‐selenium supplements) and cultured in the presence of 40 ng/mL hBMP‐4, 10 ng/mL hbFGF and 50 ng/mL VEGF. On day 4, the EB were cultured with a cocktail of hematopoietic cytokines (50 ng/mL hSCF, 20 ng/mL hFlt3L, 20 ng/mL hIL‐3 and 30 ng/mL TPO). On day 14 of culture, the differentiated cells were transferred onto FcDLL4‐coated plates and cultured in the presence of a cocktail of T lineage cytokines (10 ng/mL hFlt3L, 5 ng/mL IL‐7). After 21 days of culture, the hematopoietic cells were differentiated into CD7, CD45‐positive lymphocyte progenitor cells (iCAR‐LPC). The iCAR‐LPC were harvested and stimulated to expand with PHA to generate iCAR‐NK/ILC.

Ueda T., Kumagai A., Iriguchi S., Yasui Y., Miyasaka T., Nakagoshi K., Nakane K., Saito K., Takahashi M., Sasaki A., Yoshida S., Takasu N., Seno H., Uemura Y., Tamada K., Nakatsura T, & Kaneko S. (2020). Non–clinical efficacy, safety and stable clinical cell processing of induced pluripotent stem cell‐derived anti–glypican‐3 chimeric antigen receptor‐expressing natural killer/innate lymphoid cells. Cancer Science, 111(5), 1478-1490.

+ Open protocol

+ Expand

Directed Cardiomyocyte Differentiation from hESCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cardiomyocytes were generated from human embryonic stem cell line HES2 according to a previously published protocol[64 (link)]. Briefly, embryoid bodies were formed by plating small aggregates of human ESCs in 2 ml basic media (StemPro34, Invitrogen, containing 2 mM glutamine, 4 × 10-4 M monothioglycerol (MTG), 50 μg ml-1 ascorbic acid, Sigma, and 0.5 ng ml-1 BMP4). At days 1–4, BMP4 (10 ng/ml), bFGF(5 ng/ml) and activin A (3 ng/ml) were added for primitive-streak formation; at days 4–8, VEGF(10 ng/ml) and DKK1 (150 ng/ml), were added for mesoderm induction; after day 8, VEGF (10 ng/ml), DKK1 (150 ng/ml), and bFGF (5 ng/ml) were added for cardiac specification. Cultures were maintained in a 5% CO2/5% O2/90% N2 environment for the first 10–12 days and transferred to a 5% CO2/air environment for culture maintenance.

Zhao Y., Godier-Furnemont A., Bax N.A., Bouten C.V., Brown L.M., Fine B, & Vunjak-Novakovic G. (2022). Changes in extracellular matrix in failing human non-ischemic and ischemic hearts with mechanical unloading. Journal of molecular and cellular cardiology, 166, 137-151.

+ 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?

Revolutionizing how scientists
search and build protocols!