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Review
. 2018 Dec 7;8:582.
doi: 10.3389/fonc.2018.00582. eCollection 2018.

Technical Advances in Single-Cell RNA Sequencing and Applications in Normal and Malignant Hematopoiesis

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Free PMC article
Review

Technical Advances in Single-Cell RNA Sequencing and Applications in Normal and Malignant Hematopoiesis

Xiang-Tao Huang et al. Front Oncol. .
Free PMC article

Abstract

Single-cell RNA sequencing (scRNA-seq) has been tremendously developed in the past decade owing to overcoming challenges associated with isolation of massive quantities of single cells. Previously, cell heterogeneity and low quantities of available biological material posed significant difficulties to scRNA-seq. Cell-to-cell variation and heterogeneity are fundamental and intrinsic characteristics of normal and malignant hematopoietic cells; this heterogeneity has often been ignored in omics studies. The application of scRNA-seq has profoundly changed our comprehension of many biological phenomena, including organ development and carcinogenesis. Hematopoiesis, is actually a maturation process for more than ten distinct blood and immune cells, and is thought to be critically involved in hematological homeostasis and in sustaining the physiological functions. However, aberrant hematopoiesis directly leads to hematological malignancy, and a deeper understanding of malignant hematopoiesis will provide deeper insights into diagnosis and prognosis for patients with hematological malignancies. Here, we aim to review the recent technical progress and future prospects for scRNA-seq, as applied in physiological and malignant hematopoiesis, in efforts to further understand the hematopoietic hierarchy and to illuminate personalized therapy and precision medicine approaches used in the clinical treatment of hematological malignancies.

Keywords: hematological malignancy; hematopoietic hierarchy; malignant hematopoiesis; normal hematopoiesis; single-cell RNA sequencing.

Figures

Figure 1
Figure 1
Different scRNA-seq technologies and frontier cDNA library construction platforms. (A) Chronological chart of existing scRNA-seq technologies. (B) Workflow of scRNA-seq based on 10X Genomics platform. Briefly, suspended cells and Single-Cell 3'-reagents are added into the microfluidics chip. Subsequently, together with barcoded primer gel beads and oil-surfactant solution, GEMs are formed in a “double cross” way. Single GEMs are collected in the GEMs outlet. Next, reverse transcription is initiated inside each GEM. Afterwards, GEMs are broken and barcoded cDNAs are pooled for amplification and library construction.
Figure 2
Figure 2
Normal hematopoietic cells and malignant hematopoiesis have been studied by single cell RNA-seq. (A) Single cell RNA-seq applied to hematopoietic cells, such as HSC, DCs, Mono, and T cells. Other hematopoietic cells, like CMP, GMP, MEP, LMPP, CLP, Mega, Ery, Eos, Neu, B cells, and NK cells, are also expected to be researched. (B) Single cell RNA-seq has been applied in CML, CLL, and MM. Single cell RNA-seq is expected to be applied to other hematopoietic malignancies as well, including AML, ALL, MPN, HL, NHL, and MDS. Green ovals show normal hematopoietic cells and malignant hematopoiesis that have already been studied by single cell RNA-seq, and red ovals show hematopoietic cells and malignant hematopoiesis that have yet to be evaluated by scRNA-seq. Gray oval represents other hematopoietic malignancies apart from those listed above.

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