Comparing the Osteogenic Potential and Bone Regeneration Capacities of Dedifferentiated Fat Cells and Adipose-Derived Stem Cells In Vitro and In Vivo: Application of DFAT Cells Isolated by a Mesh Method

Background: We investigated and compared the osteogenic potential and bone regeneration capacities of dedifferentiated fat cells (DFAT cells) and adipose-derived stem cells (ASCs). Method: We isolated DFAT cells and ASCs from GFP mice. DFAT cells were established by a new culture method using a mesh...

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Autores principales:	Kiyofumi Takabatake, Masakazu Matsubara, Eiki Yamachika, Yuki Fujita, Yuki Arimura, Kazuki Nakatsuji, Keisuke Nakano, Histoshi Nagatsuka, Seiji Iida
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	dedifferentiated fat cells (DFAT cells) adipose-derived stem cells (ASCs) bone regeneration mesh culture method Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/106090eb8a6d4807a203bde31619cee0
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spelling	oai:doaj.org-article:106090eb8a6d4807a203bde31619cee02021-11-25T17:56:15ZComparing the Osteogenic Potential and Bone Regeneration Capacities of Dedifferentiated Fat Cells and Adipose-Derived Stem Cells In Vitro and In Vivo: Application of DFAT Cells Isolated by a Mesh Method10.3390/ijms2222123921422-00671661-6596https://doaj.org/article/106090eb8a6d4807a203bde31619cee02021-11-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/22/12392https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Background: We investigated and compared the osteogenic potential and bone regeneration capacities of dedifferentiated fat cells (DFAT cells) and adipose-derived stem cells (ASCs). Method: We isolated DFAT cells and ASCs from GFP mice. DFAT cells were established by a new culture method using a mesh culture instead of a ceiling culture. The isolated DFAT cells and ASCs were incubated in osteogenic medium, then alizarin red staining, alkaline phosphatase (ALP) assays, and RT-PCR (for RUNX2, osteopontin, DLX5, osterix, and osteocalcin) were performed to evaluate the osteoblastic differentiation ability of both cell types in vitro. In vivo, the DFAT cells and ASCs were incubated in osteogenic medium for four weeks and seeded on collagen composite scaffolds, then implanted subcutaneously into the backs of mice. We then performed hematoxylin and eosin staining and immunostaining for GFP and osteocalcin. Results: The alizarin red-stained areas in DFAT cells showed weak calcification ability at two weeks, but high calcification ability at three weeks, similar to ASCs. The ALP levels of ASCs increased earlier than in DFAT cells and showed a significant difference (<i>p</i> < 0.05) at 6 and 9 days. The ALP levels of DFATs were higher than those of ASCs after 12 days. The expression levels of osteoblast marker genes (osterix and osteocalcin) of DFAT cells and ASCs were higher after osteogenic differentiation culture. Conclusion: DFAT cells are easily isolated from a small amount of adipose tissue and are readily expanded with high purity; thus, DFAT cells are applicable to many tissue-engineering strategies and cell-based therapies.Kiyofumi TakabatakeMasakazu MatsubaraEiki YamachikaYuki FujitaYuki ArimuraKazuki NakatsujiKeisuke NakanoHistoshi NagatsukaSeiji IidaMDPI AGarticlededifferentiated fat cells (DFAT cells)adipose-derived stem cells (ASCs)bone regenerationmesh culture methodBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 12392, p 12392 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	dedifferentiated fat cells (DFAT cells) adipose-derived stem cells (ASCs) bone regeneration mesh culture method Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	dedifferentiated fat cells (DFAT cells) adipose-derived stem cells (ASCs) bone regeneration mesh culture method Biology (General) QH301-705.5 Chemistry QD1-999 Kiyofumi Takabatake Masakazu Matsubara Eiki Yamachika Yuki Fujita Yuki Arimura Kazuki Nakatsuji Keisuke Nakano Histoshi Nagatsuka Seiji Iida Comparing the Osteogenic Potential and Bone Regeneration Capacities of Dedifferentiated Fat Cells and Adipose-Derived Stem Cells In Vitro and In Vivo: Application of DFAT Cells Isolated by a Mesh Method
description	Background: We investigated and compared the osteogenic potential and bone regeneration capacities of dedifferentiated fat cells (DFAT cells) and adipose-derived stem cells (ASCs). Method: We isolated DFAT cells and ASCs from GFP mice. DFAT cells were established by a new culture method using a mesh culture instead of a ceiling culture. The isolated DFAT cells and ASCs were incubated in osteogenic medium, then alizarin red staining, alkaline phosphatase (ALP) assays, and RT-PCR (for RUNX2, osteopontin, DLX5, osterix, and osteocalcin) were performed to evaluate the osteoblastic differentiation ability of both cell types in vitro. In vivo, the DFAT cells and ASCs were incubated in osteogenic medium for four weeks and seeded on collagen composite scaffolds, then implanted subcutaneously into the backs of mice. We then performed hematoxylin and eosin staining and immunostaining for GFP and osteocalcin. Results: The alizarin red-stained areas in DFAT cells showed weak calcification ability at two weeks, but high calcification ability at three weeks, similar to ASCs. The ALP levels of ASCs increased earlier than in DFAT cells and showed a significant difference (<i>p</i> < 0.05) at 6 and 9 days. The ALP levels of DFATs were higher than those of ASCs after 12 days. The expression levels of osteoblast marker genes (osterix and osteocalcin) of DFAT cells and ASCs were higher after osteogenic differentiation culture. Conclusion: DFAT cells are easily isolated from a small amount of adipose tissue and are readily expanded with high purity; thus, DFAT cells are applicable to many tissue-engineering strategies and cell-based therapies.
format	article
author	Kiyofumi Takabatake Masakazu Matsubara Eiki Yamachika Yuki Fujita Yuki Arimura Kazuki Nakatsuji Keisuke Nakano Histoshi Nagatsuka Seiji Iida
author_facet	Kiyofumi Takabatake Masakazu Matsubara Eiki Yamachika Yuki Fujita Yuki Arimura Kazuki Nakatsuji Keisuke Nakano Histoshi Nagatsuka Seiji Iida
author_sort	Kiyofumi Takabatake
title	Comparing the Osteogenic Potential and Bone Regeneration Capacities of Dedifferentiated Fat Cells and Adipose-Derived Stem Cells In Vitro and In Vivo: Application of DFAT Cells Isolated by a Mesh Method
title_short	Comparing the Osteogenic Potential and Bone Regeneration Capacities of Dedifferentiated Fat Cells and Adipose-Derived Stem Cells In Vitro and In Vivo: Application of DFAT Cells Isolated by a Mesh Method
title_full	Comparing the Osteogenic Potential and Bone Regeneration Capacities of Dedifferentiated Fat Cells and Adipose-Derived Stem Cells In Vitro and In Vivo: Application of DFAT Cells Isolated by a Mesh Method
title_fullStr	Comparing the Osteogenic Potential and Bone Regeneration Capacities of Dedifferentiated Fat Cells and Adipose-Derived Stem Cells In Vitro and In Vivo: Application of DFAT Cells Isolated by a Mesh Method
title_full_unstemmed	Comparing the Osteogenic Potential and Bone Regeneration Capacities of Dedifferentiated Fat Cells and Adipose-Derived Stem Cells In Vitro and In Vivo: Application of DFAT Cells Isolated by a Mesh Method
title_sort	comparing the osteogenic potential and bone regeneration capacities of dedifferentiated fat cells and adipose-derived stem cells in vitro and in vivo: application of dfat cells isolated by a mesh method
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/106090eb8a6d4807a203bde31619cee0
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Comparing the Osteogenic Potential and Bone Regeneration Capacities of Dedifferentiated Fat Cells and Adipose-Derived Stem Cells In Vitro and In Vivo: Application of DFAT Cells Isolated by a Mesh Method

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