Single-cell RNA sequencing deconvolutes the in vivo heterogeneity of human bone marrow-derived mesenchymal stem cells

• Published in: International journal of biological sciences Vol. 17; no. 15; pp. 4192 - 4206
• Main Authors: Wang, Zun, Li, Xiaohua, Yang, Junxiao, Gong, Yun, Zhang, Huixi, Qiu, Xiang, Liu, Ying, Zhou, Cui, Chen, Yu, Greenbaum, Jonathan, Cheng, Liang, Hu, Yihe, Xie, Jie, Yang, Xucheng, Li, Yusheng, Schiller, Martin R., Chen, Yiping, Tan, Lijun, Tang, Si-Yuan, Shen, Hui, Xiao, Hong-Mei, Deng, Hong-Wen
• Format: Journal Article
• Language: English
• Published: Australia Ivyspring International Publisher Pty Ltd 01.01.2021; Ivyspring International Publisher
• Subjects: Adipocytes; Aged; Aged, 80 and over; Angiogenesis; Animals; Annotations; Antibodies; Antigens; Biomarkers; Biomedical materials; Blood diseases; Bone Density; Bone marrow; Bone Marrow Cells - classification; Bone Marrow Cells - metabolism; Bone turnover; Cartilage; CD56 Antigen - genetics; CD56 Antigen - metabolism; Cell Differentiation; Chondrocytes; Chondrocytes - physiology; Cloning; Cluster Analysis; Codification; Differentiation; Female; Fibroblasts; Gene expression; Gene Expression Regulation; Gene sequencing; Glial stem cells; Hemopoiesis; Heterogeneity; Homeostasis; Humans; In vivo methods and tests; Joint replacement surgery; Laboratories; Leukocytes (mononuclear); Male; Mesenchymal stem cells; Mesenchymal Stem Cells - metabolism; Mice; Mice, Inbred C57BL; Muscle Development; Myocytes; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - metabolism; Osteoblasts; Osteogenesis; Osteoprogenitor cells; Population; Precursors; Receptors, Nerve Growth Factor - genetics; Receptors, Nerve Growth Factor - metabolism; Research Paper; Ribonucleic acid; RNA; Sequence Analysis, RNA; Single-Cell Analysis - methods; Stem cells; Stromal cells; Subpopulations; Transcription; Transcriptomes; Vertebrae; United States > US; mesenchymal stem cell (MSC); adipogenesis; single-cell RNA sequencing (scRNA-seq); chondrogenesis; osteogenesis; bone marrow
• ISSN: 1449-2288; 1449-2288
• DOI: 10.7150/ijbs.61950

Bone marrow-derived mesenchymal stem cells (BM-MSCs) are multipotent stromal cells that have a critical role in the maintenance of skeletal tissues such as bone, cartilage, and the fat in bone marrow. In addition to providing microenvironmental support for hematopoietic processes, BM-MSCs can differentiate into various mesodermal lineages including osteoblast/osteocyte, chondrocyte, and adipocyte that are crucial for bone metabolism. While BM-MSCs have high cell-to-cell heterogeneity in gene expression, the cell subtypes that contribute to this heterogeneity in humans have not been characterized. To investigate the transcriptional diversity of BM-MSCs, we applied single-cell RNA sequencing (scRNA-seq) on freshly isolated CD271 BM-derived mononuclear cells (BM-MNCs) from two human subjects. We successfully identified LEPR CD45 BM-MSCs within the CD271 BM-MNC population, and further codified the BM-MSCs into distinct subpopulations corresponding to the osteogenic, chondrogenic, and adipogenic differentiation trajectories, as well as terminal-stage quiescent cells. Biological functional annotations of the transcriptomes suggest that osteoblast precursors induce angiogenesis coupled with osteogenesis, and chondrocyte precursors have the potential to differentiate into myocytes. We also discovered transcripts for several clusters of differentiation (CD) markers that were either highly expressed (e.g., CD167b, CD91, CD130 and CD118) or absent (e.g., CD74, CD217, CD148 and CD68) in BM-MSCs, representing potential novel markers for human BM-MSC purification. This study is the first systematic dissection of human BM-MSCs cell subtypes at the single-cell resolution, revealing an insight into the extent of their cellular heterogeneity and roles in maintaining bone homeostasis.