Single Cell Biology

Abstract

Use of Media Conditioning for Robust Single Cell Sorting of Induced Pluripotent Stem Cells

Christophe Michel Raynaud* and Sharefa Al-Mannai

Human Pluripotent Stem Cells (hPSCs) constitute a unique tool for disease modeling and functional genomics. Such applications require the generation of cell clones established by repetitive cycles of tissue growth, dissociation, and re-seeding. These cloning cycles are time-consuming and increase the likelihood of genetic instability. We defined feeder-free culture conditions on Matrigel in Stem-Flex medium to establish a single-cell cloning workflow using Fluorescent Activated Cell Sorting (FACS). The workflow relies on the Stem-Flex medium supplemented with a conditioned medium and allows the efficient cloning of newly reprogramed and established human induced pluripotent stem cells lines. This workflow is an efficient and cost-effective method for the rapid derivation of stem cell clones after reprogramming or gene editing.

Published Date: 2023-11-04; Received Date: 2023-10-02