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

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.

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

“…In terms of differentiation capacity, heparin has been shown to stimulate OB differentiation of stem cells ( Ling et al, 2010 ), while our results did not show any profound differences between heparin- and EDTA-isolated hBMSCs in terms of OB and AD differentiation. Similar results were reported in the study by Roger et al (2020) and Laner-Plamberger et al (2019) using a different source of human stem cells. Previous in vitro studies using heparin as a supplement in the expansion media provided promising results to improve the longevity of MSCs and reduce senescence during passaging of the cells ( Ling et al, 2016 ; Samsonraj et al, 2017 ; Wijesinghe et al, 2017 ).…”

The Choice of Anticoagulant Influences the Characteristics of Bone Marrow Aspirate Concentrate and Mesenchymal Stem Cell Bioactivity In Vitro

“…The objective of the present study was to assess innate differences in BMC preparations containing SC or HS, both of which are used for bone marrow aspiration [21,25]. Our study evaluated two separate concentrations of HS, where BMA was collected into HS solutions resulting in clinically relevant final concentrations of HS at 1,000 U/mL and 100 U/mL of BMA [20,21], representing Series 1 and Series 2, respectively.…”

“…The aim of the present in vitro study was to determine whether SC and HS differentially influence BMC quality, using TNC, viability, and CFU-f as metrics, along with cytokine quantification in BMC in vitro cultures to assess potential differences in cell expression profiles resulting from the BMC processing in each anticoagulant. Donor-matched BMAs were collected using clinically relevant doses of SC (15% final v:/v:) [18] and HS (1,000 U/mL and 100 U/mL final v:/v:) [19][20][21]. Here, we find that the brief exposure of the BMAs to each respective anticoagulant during the processing to BMC results in distinct differences in the quality of BMC products.…”

Impact of the Process Variables on the Yield of Mesenchymal Stromal Cells from Bone Marrow Aspirate Concentrate

“…A few studies have shown that mere rinsing of the bone marrow needle and syringe with heparin before bone marrow harvesting was sufficient [ 102 ], whereas a few studies stated the usage of 100 IU of heparin per ml of bone marrow to achieve sufficient anticoagulation [ 103 ]. Moreover, the published literature has demonstrated convincing evidence on heparin, although it belongs to bio-active substances which can potentiate the biological effects of protein binding and the sustained release of growth factors and cytokines [ 104 , 105 ].…”