Isolation of Mesenchymal Stem Cells from Bone Marrow Aspirate

Chippendale, Thomas W. E.; Coopman, Karen; Rafiq, Qasim A.; Hewitt, Christopher J.; Haj, Alicia J. El

doi:10.1016/b978-0-08-088504-9.00523-7

Cited by 3 publications

(4 citation statements)

References 73 publications

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“…Cells are finally recovered by enzymatic detachment from the magnetic beads [ purification method is flow cytometry using FACS instruments, based on the positive identification of cell surface markers expressed by MSCs. However, the lack of knowledge of unique markers for different cell types is an important limitation for this method, together with the possibility of cell contamination during the sorting phases and physical stress caused by probes, laser damage and osmotic stress on cells [145]. Both techniques are based on adherent cell purification and their detachment from the plastic surface is a critical step, considering that enzymatic dissociation can lead to proteolytic damage of cell surface proteins.…”

Section: Isolation Procedures Mattermentioning

confidence: 99%

Challenges in Clinical Development of Mesenchymal Stromal/Stem Cells: Concise Review

Mastrolia

Foppiani

Murgia

et al. 2019

Stem Cells Translational Medicine

239

182

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Summary Identified 50 years ago, mesenchymal stromal/stem cells (MSCs) immediately generated a substantial interest among the scientific community because of their differentiation plasticity and hematopoietic supportive function. Early investigations provided evidence of a relatively low engraftment rate and a transient benefit for challenging congenital and acquired diseases. The reasons for these poor therapeutic benefits forced the entire field to reconsider MSC mechanisms of action together with their ex vivo manipulation procedures. This phase resulted in advances in MSCs processing and the hypothesis that MSC‐tissue supportive functions may be prevailing their differentiation plasticity, broadening the spectrum of MSCs therapeutic potential far beyond their lineage‐restricted commitments. Consequently, an increasing number of studies have been conducted for a variety of clinical indications, revealing additional challenges and suggesting that MSCs are still lagging behind for a solid clinical translation. For this reason, our aim was to dissect the current challenges in the development of still promising cell types that, after more than half a century, still need to reach their maturity. Stem Cells Translational Medicine 2019;8:1135–1148

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Section: Isolation Procedures Mattermentioning

confidence: 99%

Challenges in Clinical Development of Mesenchymal Stromal/Stem Cells: Concise Review

Mastrolia

Foppiani

Murgia

et al. 2019

Stem Cells Translational Medicine

239

182

View full text Add to dashboard Cite

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“…The fine comparison mesh has m = 13 758 elements. We use the third mesh which has n = 4202 elements as the percentage error is less than or equal to 1.59% (as shown in figures [15][16][17]. We note that we restrict our study to values of g 0:1 since the coupling of the fluid shear to the moving boundary makes the problem stiff, hence we restrict gamma to ensure reasonable convergence (poor convergence shown in figure 18 forg ¼ 0:5).…”

Section: Appendix a Full Governing Equationsmentioning

confidence: 99%

“…Bone marrow-derived ovine MSCs (oMSCs) (P3-5), isolated using the protocol in [17], were used throughout this study. Cells were cultured in expansion media consisting of Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 1% L-glutamine and 1% penicillin-streptomycin and incubated at 37°C with 5% CO 2 until 90% confluent.…”

Section: Expansion and Mnp Labelling Of Ovine Mesenchymal Stem Cellsmentioning

confidence: 99%

Experimental and mathematical modelling of magnetically labelled mesenchymal stromal cell delivery

Yeo

Markides

Schade

et al. 2021

J. R. Soc. Interface.

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A key challenge for stem cell therapies is the delivery of therapeutic cells to the repair site. Magnetic targeting has been proposed as a platform for defining clinical sites of delivery more effectively. In this paper, we use a combined in vitro experimental and mathematical modelling approach to explore the magnetic targeting of mesenchymal stromal cells (MSCs) labelled with magnetic nanoparticles using an external magnet. This study aims to (i) demonstrate the potential of magnetic tagging for MSC delivery, (ii) examine the effect of red blood cells (RBCs) on MSC capture efficacy and (iii) highlight how mathematical models can provide both insight into mechanics of therapy and predictions about cell targeting in vivo. In vitro MSCs are cultured with magnetic nanoparticles and circulated with RBCs over an external magnet. Cell capture efficacy is measured for varying magnetic field strengths and RBC percentages. We use a 2D continuum mathematical model to represent the flow of magnetically tagged MSCs with RBCs. Numerical simulations demonstrate qualitative agreement with experimental results showing better capture with stronger magnetic fields and lower levels of RBCs. We additionally exploit the mathematical model to make hypotheses about the role of extravasation and identify future in vitro experiments to quantify this effect.

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“…The greatest challenge for these methods of isolation remains however to be the lack of an exclusive marker of identifying MSCs [ 181 ]. Further limitations include the probability for cell contamination during sorting procedures, physical stresses exerted on the cells [ 208 ], and the dependence on adherent cell purification, where the use of enzymes for cell detachment can cause proteolytic damage to cell surface proteins [ 61 ]. Some of those concerns were postulated to be overcome with the development of the CliniMACS Cell Isolation System, a device that is currently clinically approved and takes advantage of conjugating colloidal suspension of superparamagnetic microbeads to a monoclonal anti-human antibody that is capable of binding to its antigen in bone marrow, umbilical cord blood products, and peripheral blood in a sterile GMP system [ 209 ].…”

Section: Mscs' Isolation Proceduresmentioning

confidence: 99%

Mesenchymal Stem/Progenitor Cells: The Prospect of Human Clinical Translation

Rady

Abbass

El-Rashidy

et al. 2020

Stem Cells International

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Mesenchymal stem/progenitor cells (MSCs) are key players in regenerative medicine, relying principally on their differentiation/regeneration potential, immunomodulatory properties, paracrine effects, and potent homing ability with minimal if any ethical concerns. Even though multiple preclinical and clinical studies have demonstrated remarkable properties for MSCs, the clinical applicability of MSC-based therapies is still questionable. Several challenges exist that critically hinder a successful clinical translation of MSC-based therapies, including but not limited to heterogeneity of their populations, variability in their quality and quantity, donor-related factors, discrepancies in protocols for isolation, in vitro expansion and premodification, and variability in methods of cell delivery, dosing, and cell homing. Alterations of MSC viability, proliferation, properties, and/or function are also affected by various drugs and chemicals. Moreover, significant safety concerns exist due to possible teratogenic/neoplastic potential and transmission of infectious diseases. Through the current review, we aim to highlight the major challenges facing MSCs’ human clinical translation and shed light on the undergoing strategies to overcome them.

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Isolation of Mesenchymal Stem Cells from Bone Marrow Aspirate

Cited by 3 publications

References 73 publications

Challenges in Clinical Development of Mesenchymal Stromal/Stem Cells: Concise Review

Challenges in Clinical Development of Mesenchymal Stromal/Stem Cells: Concise Review

Experimental and mathematical modelling of magnetically labelled mesenchymal stromal cell delivery

Mesenchymal Stem/Progenitor Cells: The Prospect of Human Clinical Translation

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