Characterization and generation of human definitive multipotent hematopoietic stem/progenitor cells

Zhu, Yanling; Wang, Tianyu; Gu, Jiaming; Huang, Ke; Zhang, Tian; Zhang, Zhishuai; Liu, He; Tang, Jun; Mai, Yuchan; Zhang, Yanqi; Li, Yuhang; Feng, Yashu; Kang, Baoqiang; Li, Jinbing; Shan, Yongli; Chen, Qianyu; Zhang, Jian; Liang, Bing; Wang, Junwei; Gao, Minghui; Zhang, Di; Zhou, Min; Zhong, Xiaofen; Chen, Jiekai; Pei, Duanqing; Nie, Jinfu; Liu, Bing; Pan, Guangjin

doi:10.1038/s41421-020-00213-6

Cited by 22 publications

(30 citation statements)

References 40 publications

(78 reference statements)

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“…We used this approach to confirm the expression of a number of novel markers in naïve and lineage committed progenitors that we had identified by clustering and trajectory analyses. For example, we identified and confirmed CD44 as a marker for naïve progenitors which has been since reported by others in single cell sequencing experiment 69 and as an endothelial cell marker in mouse AGM. 70 We also identified CD326 (EPCAM) as a marker of haematopoietic progenitors committed to the erythroid lineage and confirmed this finding in erythroid progenitors differentiated from human haematopoietic progenitors derived from umbilical cord blood.…”

Section: Characterisation Of Hpsc-derived Hspcssupporting

confidence: 83%

Progress in the production of haematopoietic stem and progenitor cells from human pluripotent stem cells

Fidanza

Forrester

2021

Journal of Immunology and Regenerative Medicine

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Cell therapies are currently used to treat many haematological diseases. These treatments range from the long-term reconstitution of the entire haematopoietic system using the most potent haematopoietic stem cells (HSCs) to the short-term rescue with mature functional end cells such as oxygen-carrying red blood cells and cells of the immune system that can fight infection and repair tissue. Limitations in supply and the risk of transmitting infection has prompted the design of protocols to produce some of these cell types from human pluripotent stem cells (hPSCs). Although it has proven challenging to generate the most potent HSCs directly from hPSCs, significant progress has been made in the development of differentiation protocols that can successfully produce haematopoietic progenitor cells and most of the mature cell lineages. We review the key steps used in the production of haematopoietic stem and progenitor cells (HSPCs) from hPSCs and the cell surface markers and reporter strategies that have been used to define specific transitions. Most studies have relied on the use of known markers that define HSPC production in vivo but more recently single cell RNA sequencing has allowed a less biased approach to their characterisation. Transcriptional profiling has identified new markers for naïve and committed hPSC-derived HSPC populations and trajectory analyses has provided novel insights into their lineage potential. Direct comparison of in vitro- and in vivo -derived RNA single cell sequencing datasets has highlights similarities and differences between the two systems and this deeper understanding will be key to the design and the tracking of improved and more efficient differentiation protocols.

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Section: Characterisation Of Hpsc-derived Hspcssupporting

confidence: 83%

Progress in the production of haematopoietic stem and progenitor cells from human pluripotent stem cells

Fidanza

Forrester

2021

Journal of Immunology and Regenerative Medicine

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“…We found that the expression of pluripotency-related genes and transient early mesoderm markers in corrected hiPSCs was gradually decreased during the differentiation process. CD34 and CD43 are the well-known surface markers of HSCs [ 34 ]. CD34 + is used to identify hematopoietic stem cells with multilineage potential [ 35 ].…”

Section: Discussionmentioning

confidence: 99%

Genetic correction of concurrent α- and β-thalassemia patient-derived pluripotent stem cells by the CRISPR-Cas9 technology

Liu

et al. 2022

Stem Cell Res Ther

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Background Thalassemia is a genetic blood disorder characterized by decreased hemoglobin production. Severe anemia can damage organs and severe threat to life safety. Allogeneic transplantation of bone marrow-derived hematopoietic stem cell (HSCs) at present represents a promising therapeutic approach for thalassemia. However, immune rejection and lack of HLA-matched donors limited its clinical application. In recent years, human-induced pluripotent stem cells (hiPSCs) technology offers prospects for autologous cell-based therapy since it could avoid the immunological problems mentioned above. Methods In the present study, we established a new hiPSCs line derived from amniotic cells of a fetus with a homozygous β41-42 (TCTT) deletion mutation in the HBB gene and a heterozygous Westmead mutation (C > G) in the HBA2 gene. We designed a CRISPR-Cas9 to target these casual mutations and corrected them. Gene-corrected off-target analysis was performed by whole-exome capture sequencing. The corrected hiPSCs were analyzed by teratoma formation and erythroblasts differentiation assays. Results These mutations were corrected with linearized donor DNA through CRISPR/Cas9-mediated homology-directed repair. Corrections of hiPSCs were validated by sequences. The corrected hiPSCs retain normal pluripotency. Moreover, they could be differentiated into hematopoietic progenitors, which proves that they maintain the multilineage differentiation potential. Conclusions We designed sgRNAs and demonstrated that these sgRNAs facilitating the CRISPR-Cas9 genomic editing system could be applied to correct concurrent α- and β-thalassemia in patient-derived hiPSCs. In the future, these corrected hiPSCs can be applied for autologous transplantation in patients with concurrent α- and β-thalassemia.

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“…Single-cell RNA-sequencing studies, with different objectives, have yielded descriptions of a wide range of immune cells. For example, the characterization of human hematopoietic populations at the earliest stage of hematopoiesis (i.e., CD34+ cells) could provide a basis for generating blood and immune cells for clinical purposes [ 19 ], the study of CD4+ T-cells allows the specific cell states that drive disease or treatment response to be identified [ 20 ], and the analysis of stem cell-like CD8+ memory T-cells may help in the development of immunotherapies and vaccines [ 21 ].…”

Section: Applications Of Single-cell Multi-omics In Hematological mentioning

confidence: 99%

Time to Move to the Single-Cell Level: Applications of Single-Cell Multi-Omics to Hematological Malignancies and Waldenström’s Macroglobulinemia—A Particularly Heterogeneous Lymphoma

Jiménez

2021

Cancers

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Single-cell sequencing techniques have become a powerful tool for characterizing intra-tumor heterogeneity, which has been reflected in the increasing number of studies carried out and reported. We have rigorously reviewed and compiled the information about these techniques inasmuch as they are relative to the area of hematology to provide a practical view of their potential applications. Studies show how single-cell multi-omics can overcome the limitations of bulk sequencing and be applied at all stages of tumor development, giving insights into the origin and pathogenesis of the tumors, the clonal architecture and evolution, or the mechanisms of therapy resistance. Information at the single-cell level may help resolve questions related to intra-tumor heterogeneity that have not been previously explained by other techniques. With that in mind, we review the existing knowledge about a heterogeneous lymphoma called Waldenström’s macroglobulinemia and discuss how single-cell studies may help elucidate the underlying causes of this heterogeneity.

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Characterization and generation of human definitive multipotent hematopoietic stem/progenitor cells

Cited by 22 publications

References 40 publications

Progress in the production of haematopoietic stem and progenitor cells from human pluripotent stem cells

Progress in the production of haematopoietic stem and progenitor cells from human pluripotent stem cells

Genetic correction of concurrent α- and β-thalassemia patient-derived pluripotent stem cells by the CRISPR-Cas9 technology

Time to Move to the Single-Cell Level: Applications of Single-Cell Multi-Omics to Hematological Malignancies and Waldenström’s Macroglobulinemia—A Particularly Heterogeneous Lymphoma

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