Ex vivo organ culture of adipose tissue for in situ mobilization of adipose-derived stem cells and defining the stem cell niche

In spite of the advances in the knowledge of adipose‐derived stem cells (ASCs), in situ location of ASCs and the niche component of adipose tissue (AT) remain controversial due to the lack of an appropriate culture system. Here we describe a fibrin matrix‐supported three‐dimensional (3D) organ cultu...

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Veröffentlicht in:Journal of cellular physiology 2010-09, Vol.224 (3), p.807-816
Hauptverfasser: Yang, Young-Il, Kim, Hyeong-In, Choi, Min-Young, Son, Sung-Hee, Seo, Min-Jeong, Seo, Ji-Yeon, Jang, Won-Hee, Youn, Young-Chul, Choi, Kang-Joo, Cheong, Soon-Ho, Shelby, Jane
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container_end_page 816
container_issue 3
container_start_page 807
container_title Journal of cellular physiology
container_volume 224
creator Yang, Young-Il
Kim, Hyeong-In
Choi, Min-Young
Son, Sung-Hee
Seo, Min-Jeong
Seo, Ji-Yeon
Jang, Won-Hee
Youn, Young-Chul
Choi, Kang-Joo
Cheong, Soon-Ho
Shelby, Jane
description In spite of the advances in the knowledge of adipose‐derived stem cells (ASCs), in situ location of ASCs and the niche component of adipose tissue (AT) remain controversial due to the lack of an appropriate culture system. Here we describe a fibrin matrix‐supported three‐dimensional (3D) organ culture system for AT which sustains the ASC niche and allows for in situ mobilization and expansion of ASCs in vitro. AT fragments were completely encapsulated within the fibrin matrix and cultured under dynamic condition. The use of organ culture of AT resulted in a robust outgrowth and proliferation in the fibrin matrix. The outgrown cells were successfully recovered from fibrin by urokinase treatment. These outgrown cells fulfilled the criteria of mesenchymal stem cells, adherence to plastic, multilineage differentiation, and cell surface molecule expression. In vitro label retaining assay revealed that newly divided cells during the culture resided in interstitium between adipocytes and capillary endothelial cells. These interstitial stromal cells proliferated and outgrew into the fibrin matrix. Both in situ mobilized and outgrown cells expressed CD146 and α‐smooth muscle actin (SMA), but no endothelial cell markers (CD31 and CD34). The structural integrity and spatial approximation of CD31−/CD34−/CD146+/SMA+ interstitial stromal cells, adipocytes, and capillary endothelial cells were well preserved during in vitro culture. Our results suggest that ASCs are natively associated with the capillary wall and more specifically, belong to a subset of pericytes. Furthermore, organ culture of AT within a fibrin matrix‐supported 3D environment can recapitulate the ASC niche in vitro. J. Cell. Physiol. 224: 807–816, 2010. © 2010 Wiley‐Liss, Inc.
doi_str_mv 10.1002/jcp.22188
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Here we describe a fibrin matrix‐supported three‐dimensional (3D) organ culture system for AT which sustains the ASC niche and allows for in situ mobilization and expansion of ASCs in vitro. AT fragments were completely encapsulated within the fibrin matrix and cultured under dynamic condition. The use of organ culture of AT resulted in a robust outgrowth and proliferation in the fibrin matrix. The outgrown cells were successfully recovered from fibrin by urokinase treatment. These outgrown cells fulfilled the criteria of mesenchymal stem cells, adherence to plastic, multilineage differentiation, and cell surface molecule expression. In vitro label retaining assay revealed that newly divided cells during the culture resided in interstitium between adipocytes and capillary endothelial cells. These interstitial stromal cells proliferated and outgrew into the fibrin matrix. Both in situ mobilized and outgrown cells expressed CD146 and α‐smooth muscle actin (SMA), but no endothelial cell markers (CD31 and CD34). The structural integrity and spatial approximation of CD31−/CD34−/CD146+/SMA+ interstitial stromal cells, adipocytes, and capillary endothelial cells were well preserved during in vitro culture. Our results suggest that ASCs are natively associated with the capillary wall and more specifically, belong to a subset of pericytes. Furthermore, organ culture of AT within a fibrin matrix‐supported 3D environment can recapitulate the ASC niche in vitro. J. Cell. 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Cell. Physiol</addtitle><description>In spite of the advances in the knowledge of adipose‐derived stem cells (ASCs), in situ location of ASCs and the niche component of adipose tissue (AT) remain controversial due to the lack of an appropriate culture system. Here we describe a fibrin matrix‐supported three‐dimensional (3D) organ culture system for AT which sustains the ASC niche and allows for in situ mobilization and expansion of ASCs in vitro. AT fragments were completely encapsulated within the fibrin matrix and cultured under dynamic condition. The use of organ culture of AT resulted in a robust outgrowth and proliferation in the fibrin matrix. The outgrown cells were successfully recovered from fibrin by urokinase treatment. These outgrown cells fulfilled the criteria of mesenchymal stem cells, adherence to plastic, multilineage differentiation, and cell surface molecule expression. 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subjects Adipose Tissue - cytology
Adult
Biomarkers - metabolism
Cell Differentiation - physiology
Cell Proliferation
Cells, Cultured
Fibrin - metabolism
Humans
Middle Aged
Organ Culture Techniques - methods
Phenotype
Stem Cell Niche
Stem Cells - cytology
Stem Cells - physiology
Stromal Cells - cytology
Stromal Cells - physiology
Tissue Scaffolds
title Ex vivo organ culture of adipose tissue for in situ mobilization of adipose-derived stem cells and defining the stem cell niche
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