Heparin Anticoagulant for Human Bone Marrow Does Not Influence In Vitro Performance of Human Mesenchymal Stromal Cells.

Yvonne Roger,Thilo Flörkemeier,Kirsten Elger,Anika Hamm,Laura Burmeister,Oliver Dittrich-Breiholz,Andrea Hoffmann

doi:10.3390/cells9071580

Abstract

Mesenchymal stromal cells (MSCs) are a promising cell source for tissue engineering and regenerative medicine. In our lab, we found that MSC preparations from bone marrow of many different donors had a limited capacity of in vitro differentiation into osteogenic and chondrogenic lineages—a capacity claimed to be inherent to MSCs. The current study was designed to test the hypothesis that the amount of heparin used as anticoagulant during bone marrow harvest had an inhibitory influence on the in vitro differentiation capacity of isolated MSCs. Bone marrow was obtained from the femoral cavity of twelve donors during total hip arthroplasty in the absence or presence of heparin. No coagulation was observed in the absence of heparin. The number of mononuclear cells was independent of heparin addition. Isolated MSCs were characterized by morphology, population doubling times, expression of cell surface antigens and in vitro differentiation. Results of these analyses were independent of the amount of heparin. Transcriptome analyses of cells from three randomly chosen donors and quantitative realtime PCR (qRT-PCR) analysis from cells of all donors demonstrated no clear effect of heparin on the transcriptome of the cells. This excludes heparin as a potential source of disparate results.

Highlights

Mesenchymal stromal cells (MSCs) constitute a major cell population which is used in research, translational studies, and clinical applications, and bone marrow is one major tissue source
To isolate the MSCs, the heparin-anticoagulated bone marrow was diluted with PBS and processed as described in Materials and Methods
The cell populations were defined as bone marrow-MSCs based on their plastic-adherent growth, the presence or absence of cell surface antigens determined by flow cytometry and their in vitro differentiation

Summary

Introduction

Mesenchymal stromal cells (MSCs) constitute a major cell population which is used in research, translational studies, and clinical applications, and bone marrow is one major tissue source. MSCs are largely isolated and described by minimal criteria which were defined by the International Society for. Cells 2020, 9, 1580 examination of these criteria vary largely between individual labs. In research studies bone marrow is frequently obtained from patients during diagnostic or therapeutic intervention. It is known that culture conditions including the choice of fetal calf serum and use of different proteases during passaging can have an immense effect on the performance of the cells. To start at the very beginning with tissue harvest for MSC isolation, it might be important to know how the bone marrow is harvested and treated before extraction with respect to anti-coagulation—next to additional interesting aspects like the site (iliac crest or long bones) and mode of bone marrow harvest (aspiration or retrieval from an opened bone)

Results

Discussion

Conclusion