To test their colony formation capacity (CFC), cells with presumptive hemogenic capacity were sorted from 168 h gastruloids based on surface marker expression. Sorted cells were added at serial dilutions to 1.5 ml of MethoCult (Stem Cell Technologies, H4434). The mixture of MethoCult and cells was vortexed and transferred with a blunt end needle syringe (Stem Cell Technologies, 28110) into meniscus-free SmartDish plates (6-well-plate, Stem Cell Technologies, 27370). To allow constant humidification, the 6-well plates were placed in a large 150 mm square dish (Corning, CLS431111) containing small 3.5 × 10 mm tissue culture dishes filled with water. The cultures were incubated at 37 °C, 5% CO2 for 14 days and the CFC content was automatedly imaged after 7 and 14 days by using the StemVision imaging system (Stem Cell Technologies, Version 2.0.1.0). Additionally, higher-resolution images of colonies were acquired with a PALM Microbeam microscope (Zeiss) equipped with an AxioCamICc1 color camera and 5 × and 10 × objectives.
Cls431111
Manufactured by Corning
The CLS431111 is a laboratory instrument designed for conducting scientific research and analysis. It is a high-performance device that can be used for a variety of applications within the laboratory setting. The core function of the CLS431111 is to provide accurate and reliable data collection and analysis capabilities to support the needs of scientific and research professionals.
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2 protocols using cls431111
1
Assessing Hematopoietic Potential via Colony Formation
2
Quantifying Hemogenic Potential through Colony Formation Assay
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To test their colony formation capacity (CFC), cells with presumptive hemogenic capacity were sorted from day 7 gastruloids based on surface marker expression. Sorted cells were added at serial dilutions to 1.5ml of MethoCult (Stem Cell Technologies, H4434). The mixture of MethoCult and cells was vortexed and transferred with a blunt end needle syringe (Stem Cell Technologies, 28110) into meniscus-free SmartDish plates (6-well-plate, Stem Cell Technologies, 27370). To allow constant humidification, the 6-well plates were placed in a large 150mm square dish (Corning, CLS431111) containing small 3.5x10 mm tissue culture dishes filled with water. The cultures were incubated at 37°C, 5% CO2 for 14 days and the CFC content was automatedly imaged after 7 and 14 days by using the StemVision imaging system (Stem Cell Technologies, Version 2.0.1.0). Additionally, higher-resolution images of colonies were acquired with a PALM Microbeam microscope (Zeiss) equipped with an AxioCamICc1 color camera and 5x and 10x objectives.
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