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Label-free enrichment of primary human skeletal progenitor cells using deterministic lateral displacement

Label-free enrichment of primary human skeletal progenitor cells using deterministic lateral displacement
Label-free enrichment of primary human skeletal progenitor cells using deterministic lateral displacement
Skeletal stem cells (SSCs) are present in bone marrow (BM) and offer great potential for bone regenerative therapies. However, in the absence of a unique marker, current sorting approaches remain challenging in the quest for simple strategies to deliver SSCs with consistent regeneration and differentiation capacities. Microfluidics offers the possibility to sort cells marker-free, based on intrinsic biophysical properties. Recent studies indicate that SSCs are stiffer than leukocytes and are contained within the larger cell fraction in BM. This paper describes the use of deterministic lateral displacement (DLD) to sort SSCs based on cell size and stiffness. DLD is a technology that uses arrays of micropillars to sort cells based on their diameter. Cell deformation within the device can change the cell size and affect sorting – here evidenced using human cell lines and by fractionation of expanded SSCs. Following sorting, SSCs remained viable and retained their capacity to form clonogenic cultures (CFU-F), indicative of stem cell potential. Additionally, larger BM cells showed enhanced capacity to form CFU-F. These findings support the theory that SSCs are more abundant within the larger BM cell fraction and that DLD, or other size-based approaches, could be used to provide enriched SSC populations with significant implications for stem cell research and translation to the clinic.
1473-0197
513-523
Queiros De Almeida Xavier, Jorge
5587264c-be46-428a-8022-ab147a8224dd
Holm, Stefan
a4d7e8c0-1406-4f9a-8345-ddef5db5f39a
Beech, Jason
42714d13-35e2-4b32-b4e6-e24f0665b68b
Spencer, Daniel
4affe9f6-353a-4507-8066-0180b8dc9eaf
Tegenfeldt, Jonas
90546199-26e1-441c-8fc9-8a9fe746e06f
Oreffo, Richard
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174
Queiros De Almeida Xavier, Jorge
5587264c-be46-428a-8022-ab147a8224dd
Holm, Stefan
a4d7e8c0-1406-4f9a-8345-ddef5db5f39a
Beech, Jason
42714d13-35e2-4b32-b4e6-e24f0665b68b
Spencer, Daniel
4affe9f6-353a-4507-8066-0180b8dc9eaf
Tegenfeldt, Jonas
90546199-26e1-441c-8fc9-8a9fe746e06f
Oreffo, Richard
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174

Queiros De Almeida Xavier, Jorge, Holm, Stefan, Beech, Jason, Spencer, Daniel, Tegenfeldt, Jonas, Oreffo, Richard and Morgan, Hywel (2019) Label-free enrichment of primary human skeletal progenitor cells using deterministic lateral displacement. Lab on a Chip, 19 (3), 513-523. (doi:10.1039/C8LC01154K).

Record type: Article

Abstract

Skeletal stem cells (SSCs) are present in bone marrow (BM) and offer great potential for bone regenerative therapies. However, in the absence of a unique marker, current sorting approaches remain challenging in the quest for simple strategies to deliver SSCs with consistent regeneration and differentiation capacities. Microfluidics offers the possibility to sort cells marker-free, based on intrinsic biophysical properties. Recent studies indicate that SSCs are stiffer than leukocytes and are contained within the larger cell fraction in BM. This paper describes the use of deterministic lateral displacement (DLD) to sort SSCs based on cell size and stiffness. DLD is a technology that uses arrays of micropillars to sort cells based on their diameter. Cell deformation within the device can change the cell size and affect sorting – here evidenced using human cell lines and by fractionation of expanded SSCs. Following sorting, SSCs remained viable and retained their capacity to form clonogenic cultures (CFU-F), indicative of stem cell potential. Additionally, larger BM cells showed enhanced capacity to form CFU-F. These findings support the theory that SSCs are more abundant within the larger BM cell fraction and that DLD, or other size-based approaches, could be used to provide enriched SSC populations with significant implications for stem cell research and translation to the clinic.

Text
C8LC01154K - Accepted Manuscript
Restricted to Repository staff only until 9 January 2020.
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Accepted/In Press date: 7 January 2019
e-pub ahead of print date: 9 January 2019
Published date: 7 February 2019

Identifiers

Local EPrints ID: 427441
URI: https://eprints.soton.ac.uk/id/eprint/427441
ISSN: 1473-0197
PURE UUID: 63058999-3742-4902-97d0-951e68ffdd8c
ORCID for Richard Oreffo: ORCID iD orcid.org/0000-0001-5995-6726
ORCID for Hywel Morgan: ORCID iD orcid.org/0000-0003-4850-5676

Catalogue record

Date deposited: 16 Jan 2019 17:30
Last modified: 03 Dec 2019 01:54

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Contributors

Author: Jorge Queiros De Almeida Xavier
Author: Stefan Holm
Author: Jason Beech
Author: Daniel Spencer
Author: Jonas Tegenfeldt
Author: Richard Oreffo ORCID iD
Author: Hywel Morgan ORCID iD

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