FACS reveals more Pluripotent Intervertebral Disc Progenitor Cells compared to MACS and pluriSelect

Frauchiger, D.A.; May, Rahel Deborah; Dzafo, E.; Zhang, Xingshuo; Stoyanov, J.; Bertolo, A.; Benneker, Lorin Michael; Sakai, D.; Grad, S.; Tryfonidou, M.A.; Gantenbein, Benjamin (2019). FACS reveals more Pluripotent Intervertebral Disc Progenitor Cells compared to MACS and pluriSelect. In: TERMIS European Chapter Meeting 2019 (p. 898). Davos eCMjournal: www.ecmjournal.org

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INTRODUCTION: Nucleus Pulposus Progenitor Cells (NPPCs), positive for the angiopoietin-1 receptor (Tie2), were demonstrated in human, mouse, canine and bovine NP tissue [1,2,3]. Tie2+ NPPCs possess a multi-lineage differentiation potential, and regeneration potential is attributed to them. However, the isolation of Tie2+ NPPCs can be cumbersome. Hence, three isolation methods were compared. METHODS: Bovine NP cells were isolated from 10-14-month-old animals. Cell sorting was performed with an antibody against Tie2 (bs-1300R, Bioss) using FACS, magnetic-activated cell sorting (MACS) and pluriSelect, a size-based sorting method. Outcomes were evaluated by cell yield of Tie2+ cells, the ability of sorted cells to form colonies and tri-lineage differentiation assays. RESULTS & DISCUSSION: FACS resulted in the highest Tie2+ cell yield (5.0 ± 4.0%) followed by MACS (1.6 ± 2.9%) and pluriSelect (1.1 ± 1.4%). Colony forming ability did not differ between Tie2+ and Tie2- cells for any isolation method. However, Tie2+ cells obtained by MACS tended to have more colonies than FACS and pluriSelect. Osteogenic and adipogenic differentiation of Tie2+ and Tie2- cells did not result in a clear distinction for MACS and pluriSelect; Tie2+ FACS-sorted cells demonstrated superior osteogenic and adipogenic differentiation over Tie2- cells. Also for chondrogenesis, the Tie2+ FACS-sorted Tie2+ NPPCs tended to produce more proteoglycan versus Tie2- NPPCs, whereas for MACS and pluriSelect no difference was found. CONCLUSIONS: Based on the parameters tested, isolation of NPPC is possible with all three methods. However, cell yields differed widely. FACS although most invasive, appears to be the most specific sorting method for these Tie2 + cells among the tested methods as Tie2 + cells do not demonstrate osteogenic and adipogenic differentiation. As for cell yield MACS seems to reveal the most, possibly this is due to inclusion of cells expressing Tie2 less strongly.

Item Type:

Conference or Workshop Item (Poster)

Division/Institute:

?? 6F7A6E49C4DE4C679BDE034AB1478277 ??
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR)
04 Faculty of Medicine > Department of Orthopaedic, Plastic and Hand Surgery (DOPH) > Clinic of Orthopaedic Surgery
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > DBMR Forschung Mu35 > Forschungsgruppe Knochenbiologie & Orthopädische Forschung
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > DBMR Forschung Mu35 > Forschungsgruppe Knochenbiologie & Orthopädische Forschung

04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > DCR Services > Core Facility Zytometrie-Labor/FACSlab
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > DCR Services > Core Facility Live Cell Imaging (LCI)

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

May, Rahel Deborah; Zhang, Xingshuo; Benneker, Lorin Michael and Gantenbein, Benjamin

Subjects:

600 Technology > 610 Medicine & health
500 Science > 570 Life sciences; biology

Publisher:

www.ecmjournal.org

Language:

English

Submitter:

Benjamin Gantenbein

Date Deposited:

04 Sep 2019 12:28

Last Modified:

21 Nov 2019 04:49

Additional Information:

eCM Periodical, 2019, Collection 3; 2019 TERMIS EU Abstracts (page 898)

BORIS DOI:

10.7892/boris.132972

URI:

https://boris.unibe.ch/id/eprint/132972

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