Сord blood hematopoietic stem cells ex vivo enhance the bipotential commitment of adipose mesenchymal stromal progenitors

L B Buravkova; M I Ezdakova; I V Andrianova; A N Gornostaeva; P I Bobyleva; E R Andreeva

doi:10.1016/j.lfs.2020.118970

Сord blood hematopoietic stem cells ex vivo enhance the bipotential commitment of adipose mesenchymal stromal progenitors

Life Sci. 2021 Mar 1:268:118970. doi: 10.1016/j.lfs.2020.118970. Epub 2020 Dec 29.

Authors

L B Buravkova¹, M I Ezdakova², I V Andrianova², A N Gornostaeva², P I Bobyleva³, E R Andreeva²

Affiliations

¹ Institute of Biomedical Problems, Russian Academy of Sciences, Moscow, Russia; Faculty of Fundamental Medicine, Moscow State University, Moscow, Russia.
² Institute of Biomedical Problems, Russian Academy of Sciences, Moscow, Russia.
³ Institute of Biomedical Problems, Russian Academy of Sciences, Moscow, Russia; Faculty of Fundamental Medicine, Moscow State University, Moscow, Russia. Electronic address: blastoblast@gmail.com.

PMID: 33383051
DOI: 10.1016/j.lfs.2020.118970

Abstract

Aims: Stroma-dependent ex vivo expansion of hematopoietic stem progenitor cells (HSPCs) is a valid approach for cell therapy needs. Our goal was to verify whether HSPCs can affect stromal cells to optimize their functions during ex vivo expansion.

Main methods: HSPCs from cord blood (cb) were cocultured with growth-arrested adipose mesenchymal stromal cells (MSCs). Commitment-related transcriptional and secretory profiles as well as hematopoiesis-supportive activity of intact and osteo-induced MSCs were examined.

Key findings: During expansion, cbHSPCs affected the functional state of MSCs, contributing to the formation of early stromal progenitors with a bipotential osteo-adipogenic profile. This was evidenced by the upregulation of certain MSC genes of osteo- and adipodifferentiation (ALPL, RUNX2, BGLAP, CEBPA, ADIPOQ), as well as by elevated alkaline phosphatase activity and altered osteoprotein patterns. Joint paracrine profiles upon coculture were characterized by a balance of "positive" (GM-SCF) and "negative" (IP-10, MIP-1α, MCP-3) myeloid regulators, effectively supporting expansion of both committed and primitive cbHSPCs. Short-term (72 h) osteoinduction prior to coculture resulted in more pronounced shift of the bipotential transcriptomic and osteoprotein profiles. The increased proportions of late primitive CD133^-/CD34⁺cbHSPCs and unipotent CFUs suggested that cbHSPCs after expansion on osteo-MSCs were more committed versus cbHSPCs from coculture with non-differentiated MSCs.

Significance: During ex vivo expansion, cbHSPCs can drive the bipotential osteo-adipogenic commitment of MSCs, providing a specific hematopoiesis-supportive milieu. Short-term preliminary osteo-induction enhanced the development of the bipotential profile, leading to more pronounced functional polarization of cbHSPCs, which may be of interest in an applied context.

Keywords: Adipose tissue-derived mesenchymal stromal cells (MSCs); Bipotential commitment; Coculture; Cord blood hematopoietic stem and progenitor cells (cbHSPCs); Osteoproteins.

MeSH terms

Alkaline Phosphatase / metabolism
Chemokines / metabolism
Coculture Techniques
Colony-Forming Units Assay
Core Binding Factor Alpha 1 Subunit / genetics
Fetal Blood / cytology*
Gene Expression Regulation
Hematopoietic Stem Cells / cytology*
Humans
Immunophenotyping
Mesenchymal Stem Cells / cytology*
Stromal Cells / cytology

Substances

Chemokines
Core Binding Factor Alpha 1 Subunit
RUNX2 protein, human
Alkaline Phosphatase