In vitro clonal multilineage differentiation of distinct murine hematopoietic progenitor populations

Fatemeh Safi; Parashar Dhapola; Eva Erlandsson; Linda Geironson Ulfsson; Ariana S Calderón; Charlotta Böiers; Göran Karlsson

doi:10.1016/j.xpro.2022.101965

In vitro clonal multilineage differentiation of distinct murine hematopoietic progenitor populations

STAR Protoc. 2023 Mar 17;4(1):101965. doi: 10.1016/j.xpro.2022.101965. Epub 2023 Jan 11.

Authors

Fatemeh Safi¹, Parashar Dhapola¹, Eva Erlandsson¹, Linda Geironson Ulfsson¹, Ariana S Calderón¹, Charlotta Böiers¹, Göran Karlsson²

Affiliations

¹ Division of Molecular Hematology, Lund Stem Cell Center, Lund University, Lund, Sweden.
² Division of Molecular Hematology, Lund Stem Cell Center, Lund University, Lund, Sweden. Electronic address: goran.karlsson@med.lu.se.

Abstract

Here we describe an in vitro co-culture system that can differentiate hematopoietic progenitor populations to all major hematopoietic lineages at clonal level. We present both a sensitive single-cell switch-culture system as well as a less laborious alternative barcoding protocol more convenient for larger cell numbers. Importantly, generation of all lineages from single long-term hematopoietic stem cells are described, following 21 days of culture. This protocol represents an efficient tool for validation experiments for single-cell genomics data. For complete details on the use and execution of this protocol, please refer to Safi et al. (2022).¹.

Keywords: Bioinformatics; Cell Biology; Cell culture; Cell isolation; Flow Cytometry/Mass Cytometry; Genomics; Molecular Biology; Stem Cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Differentiation / genetics
Coculture Techniques
Hematopoietic Stem Cells*
Mice