Human HSCs (Lin−CD34+CD38−CD49f+) and MPPs (Lin−CD34+CD38−CD49f−) were FACS-sorted and cultured in rhTPO-free Megacult-C medium (Stem Cell Technologies) to assess colony formation of megakaryocyte progenitors, according to manufacturer’s instructions. Colonies were scored after 10–12 days using Inverted Infinity and Phase Contrast Microscope (Fisher Scientific). To assess the colony formation of myeloid and erythroid progenitors, stem and progenitor cells were plated in HSC003 (human) or HSC007 (mouse) methylcellulose medium according to the manufacturer’s recommendation (R&D Systems). Colonies were scored after 14–16 days for human and 8–12 days for mouse using Inverted Infinity and Phase Contrast Microscope (Fisher Scientific). In the serial replating assay, methylcellulose medium from primary plating was dissolved in PBS to dissociate the colonies into a single cell suspension. Subsequently, 20,000 cells from the previous plating were serially replated in 1 mL of HSC003 or HSC007 medium for human and mouse cells, respectively.
Inverted infinity
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Inverted Infinity is a laboratory microscope designed for cell culture and imaging applications. It features an inverted optical design, allowing for easy access to the sample stage and the use of tissue culture plates or dishes. The Inverted Infinity provides stable, high-quality imaging performance for a variety of microscopy techniques.
Automatically generated - may contain errors
Lab products found in correlation
Heracell 150i, by Thermo Fisher Scientific (3 mentions)
Phase contrast microscope, by Thermo Fisher Scientific (2 mentions)
Lab centrifuge, by Thermo Fisher Scientific (1 mentions)
Hemocytometer, by Hausser Scientific (1 mentions)
Hema 3, by Thermo Fisher Scientific (1 mentions)
Megacult c medium, by STEMCELL (1 mentions)
3 protocols using inverted infinity
1
Evaluating Hematopoietic Progenitor Colonies
2
Polypropylene Tube Cell Isolation Protocol
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In this work, various materials were used. These materials are 50 polypropylene conical tubes (Baxter, Toronto, ON, Canada); Sterile cell strainer: 40 μm nylon mesh (Millipore, Burlington, MA, USA); 11 μm nylon filters (Millipore, Burlington, MA, USA); 1.5 mL Eppendorf tubes (Fisherscience, Saint-Laurent, QC, Canada); 0.9% irrigation-grade sodium chloride solution (Baxter, Toronto, ON, Canada). We also employed several instruments, including an incubator (Heracell 150i, Thermo Fisher Scientific, Waltham, MA, USA); Lab Centrifuge (Sorvall ST 8, Thermo Scientific, Waltham, MA, USA); Hemocytometer (BLAUBRAND® Neubauer Millipore Sigma, Burlington, MA, USA); Inverted and phase contract Microscope (isherbrandTM Inverted Infinity, Phase contrast 10× and 20×, light splitter (100% or 20/80%), Fisher Scientific, Hampton, NH, USA).
3
Bronchoalveolar Lavage Cell Profiling
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Bronchoalveolar lavage fluid (BALF) was collected by lavaging lungs with 1.0 mL of sterile PBS. Cells were isolated by centrifugation, and total cell counts were determined using a hemocytometer (Hausser Scientific) on an Inverted Infinity and Phase Contrast Microscope (Fisher Scientific). Differential cell counts were obtained via cytospins using Hema3 stained (Fisher Scientific) total cells. Differentials were performed on a minimum of 300 cells/animal.
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