Quantification of Mouse Hematopoietic Progenitors’ Formation Using Time-lapse Microscopy and Image Analysis

Abstract

In vitro differentiation of mouse embryonic stem cells (mESCs) towards blood cells constitutes a well-established system to study the endothelial-to-hematopoietic transition (EHT) at the onset of blood development. Assessing the emergence of small non-adherent round blood cells in the culture without disturbing it is essential to evaluate the progression of EHT and also to test conditions potentially enhancing or repressing this process. Here, we describe how to quantify the formation of mouse hematopoietic progenitors during EHT in normal conditions or following over-expression of eight essential transcription factors using time-lapse microscopy and image analysis.

Single-cell transcriptomics reveals a new dynamical function of transcription factors during embryonic hematopoiesis

“…For each time point, the average and standard deviation of counts from all three spots were calculated to make graphs with Microsoft Office Excel software. A detailed explanation on how time-lapse photography and image analysis were performed can be found at Bio-protocol ( Bergiers et al, 2019 )…”