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      Comparison of Mesenchymal Stem Cell Surface Markers from Bone Marrow Aspirates and Adipose Stromal Vascular Fraction Sites

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          Abstract

          The objective of this study was to subjectively evaluate the harvest of two areas of adipose collection and three areas of bone marrow collection as potential sites for clinical harvest of adipose stromal vascular fraction (SVF) and bone marrow concentrate for clinical use by quantifying the amount of tissue harvested, subjective ease of harvest, the variation of each site, and determining the cell surface marker characteristics using commercially available antibodies. Bone marrow and adipose tissue samples were collected from 10 adult mixed breed dogs. Adipose tissue was collected from the caudal scapular region and falciform fat ligament. Bone marrow aspirates were collected from the ilium, humerus, and tibia. Tissues were weighed (adipose) or measured by volume (bone marrow), processed to isolate the SVF or bone marrow concentrate, and flow cytometry was performed to quantitate the percentage of cells that were CD90, CD44 positive, and CD45 negative. Sites and tissue types were compared using matched pairs t-test. Subjectively subcutaneous fat collection was the most difficult and large amounts of tissue dissection were necessary. Additionally the subcutaneous area yielded less than the goal amount of tissue. The bone marrow harvest ranged from 10 to 27.5 ml. Adipose tissue had the highest concentration of cells with CD90 +, CD44 +, and CD45 − markers ( P < 0.05), and bone marrow had the highest total number of these cells at harvest ( P < 0.05). Variation was high for all sites, but the adipose collection yielded more consistent results. These results describe the relative cellular components in the SVF of adipose tissue and bone marrow as defined by the biomarkers chosen. Although bone marrow yielded higher absolute cell numbers on average, adipose tissue yielded more consistent results. Fat from the falciform ligament was easily obtained with less dissection and therefore created less perceived relative patient trauma.

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          Characterization and expression analysis of mesenchymal stem cells from human bone marrow and adipose tissue.

          Ryang Hwa Lee, ByungChul Kim, IkSoo Choi … (2004)
          Human mesenchymal stem cells (MSC), that have been reported to be present in bone marrow, adipose tissues, dermis, muscles and peripheral blood, have the potential to differentiate along different lineages including those forming bone, cartilage, fat, muscle and neuron. This differentiation potential makes MSC excellent candidates for cell-based tissue engineering. In this study, we have examined phenotypes and gene expression profile of the human adipose tissue-derived stromal cells (ATSC) in the undifferentiated states, and compared with that of bone marrow stromal cells (BMSC). ATSC were enzymatically released from adipose tissues from adult human donors and were expanded in monolayer with serial passages at confluence. BMSC were harvested from the metaphysis of adult human femur. Flowcytometric analysis showed that ATSC have a marker expression that is similar to that of BMSC. ATSC expressed CD29, CD44, CD90, CD105 and were absent for HLA-DR and c-kit expression. Under appropriate culture conditions, MSC were induced to differentiate to the osteoblast, adipocyte, and chondrogenic lineages. ATSC were superior to BMSC in respect to maintenance of proliferating ability, and microarray analysis of gene expression revealed differentially expressed genes between ATSC and BMSC. The proliferating ability and differentiation potential of ATSC were variable according to the culture condition. The similarities of the phenotypes and the gene expression profiles between ATSC and BMSC could have broad implications for human tissue engineering. Copyright 2004 S. Karger AG, Basel
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            Comparison of rat mesenchymal stem cells derived from bone marrow, synovium, periosteum, adipose tissue, and muscle.

            Hideya Yoshimura, Takeshi Muneta, Akimoto Nimura … (2007)
            Mesenchymal stem cells (MSCs) are increasingly being reported as occurring in a variety of tissues. Although MSCs from human bone marrow are relatively easy to harvest, the isolation of rodent MSCs is more difficult, thereby limiting the number of experiments in vivo. To determine a suitable cell source, we isolated rat MSCs from bone marrow, synovium, periosteum, adipose, and muscle and compared their properties for yield, expansion, and multipotentiality. After two passages, the cells in each population were CD11b (-), CD45 (-), and CD90 (+). The colony number per nucleated cells derived from synovium was 100-fold higher than that for cells derived from bone marrow. With regard to expansion potential, synovium-derived cells were the highest in colony-forming efficiency, fold increase, and growth kinetics. An in vitro chondrogenesis assay demonstrated that the pellets derived from synovium were heavier, because of their greater production of cartilage matrix, than those from other tissues, indicating their superiority in chondrogenesis. Synovium-derived cells retained their chondrogenic potential after a few passages. The Oil Red-O positive colony-rate assay demonstrated higher adipogenic potential in synovium- and adipose-derived cells. Alkaline phosphatase activity was greater in periosteum- and muscle-derived cells during calcification. The yield and proliferation potential of rat MSCs from solid tissues was much better than those from bone marrow. In particular, synovium-derived cells had the greatest potential for both proliferation and chondrogenesis, indicating their usefulness for cartilage study in a rat model.
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              From heterogeneity to plasticity in adipose tissues: site-specific differences.

              Louis Casteilla, Luc Pénicaud, David Caton … (2006)
              In mammals, two types of adipose tissues are present, brown (BAT) and white (WAT). WAT itself can be divided into subcutaneous and internal fat deposits. All these tissues have been shown to present a great tissue plasticity, and recent data emphasized on the multiple differentiation potentials obtained from subcutaneous WAT. However, no study has compared the heterogeneity of stroma-vascular fraction (SVF) cells and their differentiation potentials according to the localization of the fat pad. This study clearly demonstrates that WAT and BAT present different antigenic features and differentiation potentials. WAT by contrast to BAT contains a large population of hematopoietic cells composed essentially of macrophages and hematopoietic progenitor cells. In WAT, the non-hematopoietic population is mainly composed of mesenchymal stem cell (MSC)-like but contains also a significant proportion of immature cells, whereas in BAT, the stromal cells do not present the same phenotype. Internal and subcutaneous WAT present some discrete differences in the phenotype of their cell populations. WAT derived SVF cells give rise to osteoblasts, endothelial cells, adipocytes, hematopoietic cells, and cardiomyoblasts only from inguinal cells. By contrast, BAT derived SVF cells display a reduced plasticity. Adipose tissues thus appear as complex tissues composed of different cell subsets according to the location of fat pads. Inguinal WAT appears as the most plastic adipose tissue and represents a potential and suitable source of stem cell, considering its easy sampling as a major advantage for cell therapy.
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                Author and article information

                Contributors
                Meghan O. Sullivan: URI : http://frontiersin.org/people/u/301705
                Wanda J. Gordon-Evans: URI : http://frontiersin.org/people/u/272765
                Lisa Page Fredericks: URI : http://frontiersin.org/people/u/305073
                Kristina Kiefer: URI : http://frontiersin.org/people/u/306932
                Michael G. Conzemius: URI : http://frontiersin.org/people/u/304375
                Dominique J. Griffon: URI : http://frontiersin.org/people/u/245803
                Journal
                Journal ID (nlm-ta): Front Vet Sci
                Journal ID (iso-abbrev): Front Vet Sci
                Journal ID (publisher-id): Front. Vet. Sci.
                Title: Frontiers in Veterinary Science
                Publisher: Frontiers Media S.A.
                ISSN (Electronic): 2297-1769
                Publication date (Electronic): 15 January 2016
                Publication date Collection: 2015
                Volume: 2
                Electronic Location Identifier: 82
                Affiliations
                [1] 1Angell Animal Medical Center , Boston, MA, USA
                [2] 2Wisconsin Veterinary Referral Center , Waukesha, WI, USA
                [3] 3Department of Entomology, University of Illinois at Urbana–Champaign , Urbana, IL, USA
                [4] 4St. Paul Department of Veterinary Clinical Sciences, University of Minnesota , Minneapolis, MN, USA
                [5] 5College of Veterinary Medicine, Western University of Health Sciences , Pomona, CA, USA
                Author notes

                Edited by: Fausto Cremonesi, Università degli Studi di Milano, Italy

                Reviewed by: Ruchi Sharma, Axol Biosciences, UK; Sarah Elizabeth Taylor, University of Edinburgh, UK

                *Correspondence: Dominique J. Griffon, dgriffon@ 123456westernu.edu

                Specialty section: This article was submitted to Veterinary Regenerative Medicine, a section of the journal Frontiers in Veterinary Science

                Article
                DOI: 10.3389/fvets.2015.00082
                PMC ID: 4713840
                PubMed ID: 26835460
                SO-VID: 92e9c943-757c-482f-a63b-30e3059b8786
                Copyright © Copyright © 2016 Sullivan, Gordon-Evans, Fredericks, Kiefer, Conzemius and Griffon.
                License:

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                Date received : 01 October 2015
                Date accepted : 21 December 2015
                Page count
                Figures: 4, Tables: 2, Equations: 0, References: 46, Pages: 9, Words: 7311
                Funding
                Funded by: Veterinary Centers of America
                Funded by: University of Illinois at Urbana-Champaign 10.13039/100005302
                Funded by: University of Minnesota 10.13039/100007249
                Categories
                Subject: Veterinary Science
                Subject: Original Research

                Keywords: canine,adipose,bone marrow,stromal vascular fraction,mesenchymal stem cell
                Data availability:
                Keywords: canine, adipose, bone marrow, stromal vascular fraction, mesenchymal stem cell

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