{Reference Type}: Journal Article
{Title}: A culture platform to study quiescent hematopoietic stem cells following genome editing.
{Author}: Shiroshita K;Kobayashi H;Watanuki S;Karigane D;Sorimachi Y;Fujita S;Tamaki S;Haraguchi M;Itokawa N;Aoyoama K;Koide S;Masamoto Y;Kobayashi K;Nakamura-Ishizu A;Kurokawa M;Iwama A;Okamoto S;Kataoka K;Takubo K;
{Journal}: Cell Rep Methods
{Volume}: 2
{Issue}: 12
{Year}: 12 2022 19
暂无{DOI}: 10.1016/j.crmeth.2022.100354
{Abstract}: Other than genetically engineered mice, few reliable platforms are available for the study of hematopoietic stem cell (HSC) quiescence. Here we present a platform to analyze HSC cell cycle quiescence by combining culture conditions that maintain quiescence with a CRISPR-Cas9 genome editing system optimized for HSCs. We demonstrate that preculture of HSCs enhances editing efficiency by facilitating nuclear transport of ribonucleoprotein complexes. For post-editing culture, mouse and human HSCs edited based on non-homologous end joining and cultured under low-cytokine, low-oxygen, and high-albumin conditions retain their phenotypes and quiescence better than those cultured under the proliferative conditions. Using this approach, HSCs regain quiescence even after editing by homology-directed repair. Our results show that low-cytokine culture conditions for gene-edited HSCs are a useful approach for investigating HSC quiescence ex vivo.