Log In Sign up
The largest database of trusted experimental protocols

The HTHPO is a high-throughput, high-pressure oxygen analyzer designed for industrial and laboratory applications. It is capable of accurately measuring oxygen concentrations in a variety of gas streams at high pressures up to 3,000 psi. The HTHPO provides reliable and consistent performance, allowing users to monitor oxygen levels in their processes effectively.

Automatically generated - may contain errors

Lab products found in correlation

Mil 3, by Thermo Fisher Scientific (3 mentions) Hil 6, by Thermo Fisher Scientific (2 mentions) Acetylthiocholiniodide staining, by Merck Group (2 mentions) Hil 11, by Thermo Fisher Scientific (2 mentions) Hthpo, by Roche (2 mentions) Mil 3, by Roche (2 mentions) Pen strep, by Thermo Fisher Scientific (1 mentions) Mil 3, by R&D Systems (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) β mercaptoethanol, by Merck Group (1 mentions) Inverted fluorescence microscope, by Olympus (1 mentions) X vivo 15 medium, by Lonza (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Methocult m3434, by STEMCELL (1 mentions) Methocult m3436, by STEMCELL (1 mentions) Stemspan sfem, by STEMCELL (1 mentions)

4 protocols using hthpo

1

Lineage Potential of Sorted HSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
For the evaluation of lineage potential and timing of Mk cell emergence single Vwf+BiotinHi, Vwf+BiotinLo, Vwf–BiotinHi or Vwf–BiotinLo HSCs were sorted directly into 20uL X-Vivo15 medium (Lonza) supplemented with 10% fetal bovine serum (Sigma); 1% Pen/Strep (Gibco); 10-4M β-Mercaptoethanol (Sigma); mSCF (10 ug/mL; R&D Systems); hFlt3L (10 ug/mL; Immunex); hTHPO (10 ug/mL; PeproTech); mIL-3 (5 ug/mL; R&D Systems). Cultures were evaluated every day between days 2 and 8 for morphology and GFP expression using an inverted fluorescence microscope (Olympus). Fluorescent images were processed with Fiji software (ImageJ). Presence of Mk and myeloid (GM) cells was confirmed by May-Grunwald Giemsa (MGG)-stained cytospin preparations from selected wells (after 8 days of culture).
Luis T.C., Barkas N., Carrelha J., Giustacchini A., Mazzi S., Norfo R., Wu B., Aliouat A., Guerrero J.A., Rodriguez-Meira A., Bouriez-Jones T., Macaulay I.C., Jasztal M., Zhu G., Ni H., Robson M.J., Blakely R.D., Mead A.J., Nerlov C., Ghevaert C, & Jacobsen S.E. (2023). Perivascular niche cells sense thrombocytopenia and activate hematopoietic stem cells in an IL-1 dependent manner. Nature Communications, 14, 6062.
+ Open protocol
+ Expand
2

Evaluating Erythroid and Megakaryocytic Potential

Check if the same lab product or an alternative is used in the 5 most similar protocols
For evaluation of erythroid potential from progenitor populations, cells were seeded in Methocult M3436 and myeloid potential was evaluated with Methocult M3534. Colonies were scored after 8 days of culture. CFU-Mix colonies were scored in Methocult M3434 based on colonies containing red cells after 8 days of culture. Megakaryocytic potential was evaluated using the Megacult collagen-based assay. All these media were from StemCell Technologies, Vancouver, Canada. Megakaryocyte cultures were supplemented with 10ng/ml mIL-3, 20ng/ml hIL-6 (Peprotech), 50ng/ml hIL-11 (Peprotech), 50ng/ml hThpo (Peprotech), and megakaryocytes detected using acetylthiocholiniodide staining (Sigma) according to manufacturer’s instructions after 7-8 days culture. For evaluation of Mk potential from single GE+ and GE− Lin−Sca-1+c-Kit+Flt3hi cells, these cells were sorted into X-vivo15 medium containing glutamax and gentamycin (BioWhittaker) supplemented with 10% fetal calf serum (Hyclone), 10-4M β-mercaptoethanol, 50ng/mL mSCF, 50ng/mL hFLT3 ligand (hFL; Immunex), 50ng/mL hThpo, 20ng/mL mIL-3 and 5U/ml hEPO (Roche). For each experiment, 240 cells were manually plated at 1 cell per well into 60 well Terasaki plates. Mk potential was evaluated 8 or 12 days after culture using an inverted microscope.
Drissen R., Buza-Vidas N., Woll P., Thongjuea S., Gambardella A., Giustacchini A., Mancini E., Zriwil A., Lutteropp M., Grover A., Mead A., Sitnicka E., Jacobsen S.E, & Nerlov C. (2016). Distinct myeloid progenitor differentiation pathways identified through single cell RNA sequencing. Nature immunology, 17(6), 666-676.
+ Open protocol
+ Expand
3

Hematopoietic Niche Regulation by Stromal and Endothelial Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
6,000-10,000 E12.5 FL endothelial or stromal cells were FACS sorted and co-cultured with 150 FACS sorted E12.5 FL LSK cells for 5 d at 37 ºC in 200 µl of StemSpan SFEM (StemCell Technologies) supplemented with 1% P/S, 1% L-glutamine and cytokines mIL3 (5 ngml−1, PeproTech), and hTHPO (10 ngml−1, PeproTech). Media were half-changed at day 3. Non-adherent hematopoietic cells were harvested at day 5 and cultured with Methocult M3434 (Stem Cell Technologies) for 7 d at 37 ºC. Burst forming unit-erythroid (BFU-E) colony formation was determined by 2,7-diaminofluorene (DAF, Sigma) staining. For Mmp9 inhibitor 24 (link) (10 nM, CAS 1177749-58-4, Millipore) treatment, the inhibitor or DMSO (control) were added to the co-culture from day 1 and replenished at day 3 while changing media.
5,000 MS-5 cells were cultured in 10% FCS/IMDM for 24 h at 37 ºC for prior treatment with 10mM GSK126, 10mM Mmp9 inhibitor or DMSO for 24 h. 200 FACS sorted BM LSK cells were then added to the co-culture with or without drug treatment for 72 h at 37 ºC in 200 µl of StemSpan SFEM complete medium as indicated above. Non-adherent hematopoietic cells were harvested at day 3 and cultured with Methocult M3436 (Stem Cell Technologies) for 7 d at 37 ºC. BFU-E colony formation was determined by DAF staining.
Neo W.H., Booth C.A., Azzoni E., Chi L., Delgado-Olguín P., de Bruijn M.F., Jacobsen S.E, & Mead A.J. (2018). Cell-extrinsic hematopoietic impact of Ezh2 inactivation in fetal liver endothelial cells. Blood, 131(20), 2223-2234.
+ Open protocol
+ Expand
4

Evaluating Erythroid and Megakaryocytic Potential

Check if the same lab product or an alternative is used in the 5 most similar protocols
For evaluation of erythroid potential from progenitor populations, cells were seeded in Methocult M3436 and myeloid potential was evaluated with Methocult M3534. Colonies were scored after 8 days of culture. CFU-Mix colonies were scored in Methocult M3434 based on colonies containing red cells after 8 days of culture. Megakaryocytic potential was evaluated using the Megacult collagen-based assay. All these media were from StemCell Technologies, Vancouver, Canada. Megakaryocyte cultures were supplemented with 10ng/ml mIL-3, 20ng/ml hIL-6 (Peprotech), 50ng/ml hIL-11 (Peprotech), 50ng/ml hThpo (Peprotech), and megakaryocytes detected using acetylthiocholiniodide staining (Sigma) according to manufacturer’s instructions after 7-8 days culture. For evaluation of Mk potential from single GE+ and GE− Lin−Sca-1+c-Kit+Flt3hi cells, these cells were sorted into X-vivo15 medium containing glutamax and gentamycin (BioWhittaker) supplemented with 10% fetal calf serum (Hyclone), 10-4M β-mercaptoethanol, 50ng/mL mSCF, 50ng/mL hFLT3 ligand (hFL; Immunex), 50ng/mL hThpo, 20ng/mL mIL-3 and 5U/ml hEPO (Roche). For each experiment, 240 cells were manually plated at 1 cell per well into 60 well Terasaki plates. Mk potential was evaluated 8 or 12 days after culture using an inverted microscope.
Drissen R., Buza-Vidas N., Woll P., Thongjuea S., Gambardella A., Giustacchini A., Mancini E., Zriwil A., Lutteropp M., Grover A., Mead A., Sitnicka E., Jacobsen S.E, & Nerlov C. (2016). Distinct myeloid progenitor differentiation pathways identified through single cell RNA sequencing. Nature immunology, 17(6), 666-676.
+ 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!