Figure 1. Multipotency of hematopoietic stem cells.

Observation of a mitosis in an umbilical cord blood isolated CD34+ HSC sample

A sample of human cord blood CD34+ stem/progenitor cells kindly provided by Lonza was cultured in X-VIVO^TM 15 serum-free hematopoietic cell medium, supplemented with recombinant human thrombopoietin (25ng/mL), Rlt3 ligand (25ng/mL) and stem cell factor (13ng/mL). The culture was also coated with fibronectin.

A remarkably long live cell observation (3 images/min during 15hours) was performed with Nanolive’s 3D Cell Explorer, and a cell division was captured. To date and to our knowledge, this live cell movie featuring CD34+ HSC is the first one ever obtained.

One can distinguish characteristic steps of mitosis such as chromatin condensation inside the nucleus that lead to chromosome formation, the alignment of the chromosomes in the metaphase plate and the migration of sister chromatids to opposite ends of the cells. Eventually, two daughter cells were obtained as a result of the contractile ring effect during cytokinesis (Figure 2).

Considering that any stress would trigger programmed cell death in such a sensitive cell type, the mitosis captured acquires further significance. Indeed, it is a sign of not just the homogeneity of the cell culture, but also of the fact that both the medium and freezing procedures of the sample were optimal and that Nanolive’s technology was completely non-invasive, which can also be reaffirmed by observing healthy behavior in most of the cells surrounding the dividing cell.

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