Human bone marrow stromal cells: the impact of anticoagulants on stem cell properties

0578173 - FGÚ 2024 RIV CH eng J - Journal Article
Ferenčáková, Michaela - Beňová, Andrea - Raška Jr., I. - Abaffy, Pavel - Šindelka, Radek - Džubanová, Martina - Pospíšilová, E. - Kolostová, K. - Čajka, Tomáš - Paclík, A. - Zikán, V. - Tencerová, Michaela
Human bone marrow stromal cells: the impact of anticoagulants on stem cell properties.
Frontiers in Cell and Developmental Biology. Roč. 11, 18 Sep (2023), č. článku 1255823. ISSN 2296-634X. E-ISSN 2296-634X
R&D Projects: GA ČR(CZ) GA22-12243S; GA MŠMT(CZ) LX22NPO5104; GA MZd(CZ) NU23-01-00125
Grant - others:Novo Nordisk Fonden(DK) NNF20SA0066174
Institutional support: RVO:67985823 ; RVO:86652036
Keywords : human bone marrow stromal cells * anticoagulants * cultivation * stem cell characteristics * differentiation potential
OECD category: Cell biology; Developmental biology (BTO-N)
Impact factor: 5.5, year: 2022
Method of publishing: Open access
https://doi.org/10.3389/fcell.2023.1255823

Background: Bone marrow stromal cells (BMSCs) are the source of multipotent stem cells, which are important for regenerative medicine and diagnostic purposes. The isolation of human BMSCs from the bone marrow (BM) cavity using BM aspiration applies the method with collection into tubes containing anticoagulants. Interactions with anticoagulants may affect the characteristics and composition of isolated BMSCs in the culture. Thus, we investigated how anticoagulants in isolation procedures and cultivation affect BMSC molecular characteristics.Methods: BM donors (age: 48–85 years) were recruited from the hematology clinic. BM aspirates were obtained from the iliac crest and divided into tubes coated with ethylenediaminetetraacetic acid (EDTA) or heparin anticoagulants. Isolated BMSCs were analyzed by flow cytometry and RNA-seq analysis. Further cellular and molecular characterizations of BMSCs including CFU, proliferation and differentiation assays, cytometry, bioenergetic assays, metabolomics, immunostaining, and RT-qPCR were performed.Results: The paired samples of isolated BMSCs obtained from the same patient showed increased cellular yield in heparin vs. EDTA samples, accompanied by the increased number of CFU colonies. However, no significant changes in molecular characteristics were found between heparin- and EDTA-isolated BMSCs. On the other hand, RNA-seq analysis revealed an increased expression of genes involved in nucleotide metabolism and cellular metabolism in cultivated vs. non-cultivated BMSCs regardless of the anticoagulant, while genes involved in inflammation and chromatin remodeling were decreased in cultivated vs. non-cultivated BMSCs.Conclusion: The type of anticoagulant in BMSC isolation did not have a significant impact on molecular characteristics and cellular composition, while in vitro cultivation caused the major change in the transcriptomics of BMSCs, which is important for future protocols using BMSCs in regenerative medicine and clinics.
Permanent Link: https://hdl.handle.net/11104/0347212