In vitro hematopoietic support of human umbilical cord blood CD34+ cells by
human skeletal muscle-derived pericytes/perivascular cells

doi:10.3969/j.issn.2095-4344.2065

Abstract

Abstract:

BACKGROUND: Perivascular cells have been shown to be the precursor cells of mesenchymal stem cells, which regulate the behavior of hematopoietic stem cells and support hematopoiesis through cell-to-cell contact or paracrine effects. Hematopoietic support of human skeletal muscle-derived pericytes/perivascular cells (hMD-PCs) remains to be studied.

OBJECTIVE: To identify the biological characteristics of hMD-PCs isolated from human skeletal muscle and to study their supporting effect on umbilical cord blood CD34⁺ cells in vitro.

METHODS: (1) hMD-PCs with phenotype CD146⁺CD56^-CD34^-CD144^-CD45^- were sorted from human skeletal muscle by enzymatic digestion and multiparameter fluorescence-activated cell sorting, and their biological characteristics were identified. (2) The in vitro culture system of umbilical cord blood CD34⁺ cells co-cultured with human CD146⁺ hMD-PCs (experimental group) or with human bone marrow mesenchymal stem cells (positive control group) was established. After 1, 2 and 4 weeks of co-culture, the number of cells, the colony formation ability and immunophenotype were measured and statistically analyzed.

RESULTS AND CONCLUSION: (1) CD146⁺ hMD-PCs were sorted by multiparameter fluorescence-activated cell sorting and the purity was (91.5±1.85)% (n=5). CD146⁺ hMD-PCs expressed mesenchymal surface markers CD73, CD90, CD105, CD44, and did not express hematopoietic cell and endothelial cell markers CD45, CD34, and CD31. After induced culture, CD146⁺ hMD-PCs could differentiate into osteoblasts, chondrogenesis, adipocytes and myoblasts. (2) There were no significant differences in the cell number, colony formation ability or immunophenotype (CD45⁺, CD34⁺CD33^-, CD14⁺, CD10⁺/CD19⁺) between experimental and positive control groups (P > 0.05, n=6). The number of cells in the blank control group without feeder was significantly decreased at 1 week of culture, and there was almost no cell survival at 2 weeks of culture. (3) In summary, CD146⁺ hMD-PCs, like human bone marrow mesenchymal stem cells, have hematopoietic support capacity in vitro.

Key words: human skeletal muscle-derived pericytes/perivascular cells, umbilical cord blood CD34⁺cells, human bone marrow mesenchymal stem cells, trophoderm, in vitro support

CLC Number:

Yang Xiaoping, Yang Tingting, Gu Jingjing, Zhou Rui, Xu Fei, Yang Jihui, Zheng Bo. In vitro hematopoietic support of human umbilical cord blood CD34⁺ cells by human skeletal muscle-derived pericytes/perivascular cells[J]. Chinese Journal of Tissue Engineering Research, 2020, 24(13): 2047-2054.

Figures/Tables 6

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