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Water jet-assisted lipoaspiration and Sepax cell separation system for the isolation of adipose stem cells with high adipogenic potential
Umeå University, Faculty of Medicine, Department of Integrative Medical Biology (IMB). Umeå University, Faculty of Medicine, Department of Surgical and Perioperative Sciences, Hand Surgery.
Umeå University, Faculty of Medicine, Department of Surgical and Perioperative Sciences, Hand Surgery.
Umeå University, Faculty of Medicine, Department of Integrative Medical Biology (IMB). Umeå University, Faculty of Medicine, Department of Surgical and Perioperative Sciences, Hand Surgery.
Umeå University, Faculty of Medicine, Department of Clinical Microbiology.
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2021 (English)In: Journal of Plastic, Reconstructive & Aesthetic Surgery, ISSN 1748-6815, E-ISSN 1878-0539, Vol. 74, no 10, p. 2759-2767Article in journal (Refereed) Published
Abstract [en]

Introduction: Water jet-assisted liposuction has gained popularity due to favourable fat grafting outcomes. In this study, we compared stem cells obtained from fat isolated with manual or the water jet-assisted procedure.

Methods: Liposuction of abdominal fat was performed using the two methods on each donor (n = 10). Aspirate samples were collagenase digested and the isolated cells seeded in vitro prior to proliferation, adipogenic differentiation and angiogenic activity analyses.

Results: Cells from either procedure proliferated at similar rates and exhibited a similar colony-forming ability. The cells expressed stem cell markers CD73, CD90 and CD105. In the water jet cell preparations, there were higher numbers of cells expressing CD146. Robust adipogenic differentiation was observed in cultures expanded from both manual and water jet lipoaspirates. Gene analysis showed higher expression of the adipocyte markers aP2 and GLUT4 in the adipocyte-differentiated water jet cell preparations, and ELISA indicated increased secretion of adiponectin from these cells. Both cell groups expressed vasculogenic factors and the water jet cells promoted the highest levels of in vitro angiogenesis. Given these positive results, we further characterised the water jet cells when prepared using an automated closed cell processing unit, the Sepax-2 system (Cytiva). The growth and stem cell properties of the Sepax-processed cells were similar to the standard centrifugation protocol, but there was evidence for greater adipogenic differentiation in the Sepax-processed cells.

Conclusions: Water jet lipoaspirates yield cells with high adipogenic potential and angiogenic activity, which may be beneficial for use in cell-assisted lipotransfers.

Place, publisher, year, edition, pages
Elsevier, 2021. Vol. 74, no 10, p. 2759-2767
Keywords [en]
adipogenesis, angiogenesis, differentiation, fat grafting, stem cells
National Category
Cell and Molecular Biology
Identifiers
URN: urn:nbn:se:umu:diva-183242DOI: 10.1016/j.bjps.2021.03.025ISI: 000709899500011PubMedID: 33994109Scopus ID: 2-s2.0-85105758965OAI: oai:DiVA.org:umu-183242DiVA, id: diva2:1555960
Funder
Vinnova, 2017-02130Region VästerbottenAvailable from: 2021-05-19 Created: 2021-05-19 Last updated: 2024-07-02Bibliographically approved
In thesis
1. Optimizing stem cells for reconstructive surgery
Open this publication in new window or tab >>Optimizing stem cells for reconstructive surgery
2024 (English)Doctoral thesis, comprehensive summary (Other academic)
Alternative title[sv]
Optimering av stamceller för rekonstruktiv kirurgi
Abstract [en]

Fat grafting has become an established method in plastic surgery for treating soft tissue defects. The results for survival of the fat being transplanted is unpredictable and supplementation of the graft with the Stromal Vascular Fraction (SVF) or cultures Adipose tissue-derived stem cells (ASCs) can enhance graft viability. The ASCs are a heterogenous group of cells with various cell membrane markers, and differing growth promoting and differentiation characteristics of the stem cells derived from the fat. It is of high importance when expanding cells prior to the transplantation of the cells into patients, that the culture conditions are well defined and ideally are xenofree, avoiding use of animal-derived products. Furthermore, the procedures must be safe and not increase risk for recurrence of cancer after reconstructive surgeries. This thesis explores the phenotypic properties of a selected population of ASCs, with a view to determining their suitability for transplantation into fat grafts. ASCs were isolated from SVF of human abdominal fat and CD146+ cells were selected using immunomagnetic beads. The proliferation, angiogenic and adipogenic properties were significantly higher in the CD146+ cells. Stem cells were also isolated from lipoaspirate obtained using two different liposuction methods. Waterjet lipoaspirates yielded the greatest number of CD146+ cells with high adipogenic potential and angiogenic activity. The cells could also be successfully isolated using a closed processing system. Cells were expanded in either foetal bovine serum, platelet lysate or a chemically defined xenofree (XV) medium. Cultures in XV medium proliferated the fastest, expressed the highest number of CD146+ cells, and showed the best adipogenic and angiogenic properties. To test possible ASCs interactions with cancer cells, co-cultures with MCF-7 breast cancer cells were established. Conditioned medium from co-cultures significantly increased the migration of the cancer cells but not their proliferation, and there was increased expression of Tenascin-C in these cultures. The research in this thesis work has shown more optimal ways to isolate and expand ASCs, potentially offering new therapeutic reconstructive treatment options for a variety of medical conditions.

Place, publisher, year, edition, pages
Umeå: Umeå University, 2024. p. 64
Series
Umeå University medical dissertations, ISSN 0346-6612 ; 2281
Keywords
Adipose tissue-derived stem cells, liposuction, oncological safety, xenofree growth medium
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Research subject
Human Anatomy
Identifiers
urn:nbn:se:umu:diva-220039 (URN)9789180702638 (ISBN)9789180702621 (ISBN)
Public defence
2024-02-23, Sal N410, Naturvetarhuset, Umeå, 09:00 (English)
Opponent
Supervisors
Available from: 2024-02-02 Created: 2024-01-26 Last updated: 2024-01-29Bibliographically approved

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Lauvrud, Anne ThereseGümüsçü, RojdaWiberg, RebeccaBrohlin, MariaKelk, PeymanWiberg, MikaelKingham, Paul J.

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Lauvrud, Anne ThereseGümüsçü, RojdaWiberg, RebeccaBrohlin, MariaKelk, PeymanWiberg, MikaelKingham, Paul J.
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Department of Integrative Medical Biology (IMB)Hand SurgeryDepartment of Clinical Microbiology
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Journal of Plastic, Reconstructive & Aesthetic Surgery
Cell and Molecular Biology

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