Marrow changes and reduced proliferative capacity of mesenchymal stromal cells from patients with "no-option" critical limb ischemia; observations on feasibility of the autologous approach from a clinical trial

Sara Azhari Mohamed; Aoife Duffy; Veronica McInerney; Janusz Krawczyk; Amjad Hayat; Sean Naughton; Andrew Finnerty; Miriam Holohan; Aaron Liew; Muhammad Tubassam; Stewart Redmond Walsh; Timothy O'Brien; Linda Howard

doi:10.1016/j.jcyt.2022.07.002

Marrow changes and reduced proliferative capacity of mesenchymal stromal cells from patients with "no-option" critical limb ischemia; observations on feasibility of the autologous approach from a clinical trial

Cytotherapy. 2022 Dec;24(12):1259-1267. doi: 10.1016/j.jcyt.2022.07.002. Epub 2022 Aug 20.

Authors

Affiliations

¹ Regenerative Medicine Institute, National University of Ireland Galway, Galway, Ireland.
² Centre for Cell Manufacturing Ireland, National University of Ireland Galway, Galway, Ireland.
³ HRB Clinical Research Facility, National University of Ireland Galway, Galway, Ireland.
⁴ Galway Blood and Tissue Establishment, National University of Ireland Galway, Galway, Ireland.
⁵ School of Medicine, National University of Ireland Galway, Galway, Ireland.
⁶ Department of Vascular Surgery, University Hospital Galway, Galway, Ireland.
⁷ School of Medicine, National University of Ireland Galway, Galway, Ireland; Department of Vascular Surgery, University Hospital Galway, Galway, Ireland.
⁸ Regenerative Medicine Institute, National University of Ireland Galway, Galway, Ireland; Centre for Cell Manufacturing Ireland, National University of Ireland Galway, Galway, Ireland; School of Medicine, National University of Ireland Galway, Galway, Ireland.
⁹ Regenerative Medicine Institute, National University of Ireland Galway, Galway, Ireland. Electronic address: linda.howard@nuigalway.ie.

PMID: 35999133
DOI: 10.1016/j.jcyt.2022.07.002

Abstract

Background aims: Approximately 1 in 3 patients with critical limb ischemia (CLI) are not suitable for surgical or endovascular revascularization. Those "no-option" patients are at high risk of amputation and death. Autologous bone marrow mesenchymal stromal cells (MSCs) may provide a limb salvage option. In this study, bone marrow characteristics and expansion potentials of CLI-derived MSCs produced during a phase 1b clinical trial were compared with young healthy donor MSCs to determine the feasibility of an autologous approach. Cells were produced under Good Manufacturing Practice conditions and underwent appropriate release testing.

Methods: Five bone marrow aspirates derived from patients with CLI were compared with six young healthy donor marrows in terms of number of colony-forming units-fibroblast (CFUF) and mononuclear cells. The mean population doubling times and final cell yields were used to evaluate expansion potential. The effect of increasing the volume of marrow on the CFUF count and final cell yield was evaluated by comparing 5 CLI-derived MSCs batches produced from a targeted 30 mL of marrow aspirate to five batches produced from a targeted 100 mL of marrow.

Results: CLI-derived marrow aspirate showed significantly lower numbers of mononuclear cells with no difference in the number of CFUFs when compared with healthy donors' marrow aspirate. CLI-derived MSCs showed a significantly longer population doubling time and reduced final cell yield compared with young healthy donors' MSCs. The poor growth kinetics of CLI MSCs were not mitigated by increasing the bone marrow aspirate from 30 to 100 mL.

Conclusions: In addition to the previously reported karyotype abnormalities in MSCs isolated from patients with CLI, but not in cells from healthy donors, the feasibility of autologous transplantation of bone marrow MSCs for patients with no-option CLI is further limited by the increased expansion time and the reduced cell yield.

Keywords: Abnormal karyotype; Autologous approach; Clinical trial; Critical limb ischemia; Mesenchymal stromal/stem cells; Proliferative capacity.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Bone Marrow*
Chronic Limb-Threatening Ischemia
Feasibility Studies
Humans
Mesenchymal Stem Cells*
Transplantation, Autologous