Characterization of canine adipose tissue-derived mesenchymal stem cells immortalized by SV40-T retrovirus for therapeutic use

Ana Patricia Ayala-Cuellar; Cho Won Kim; Kyung A. Hwang; Ji Houn Kang; Gabsang Lee; Jaejin Cho; Kyung Chul Choi

doi:10.1002/jcp.28338

Characterization of canine adipose tissue-derived mesenchymal stem cells immortalized by SV40-T retrovirus for therapeutic use

Ana Patricia Ayala-Cuellar, Cho Won Kim, Kyung A. Hwang, Ji Houn Kang, Gabsang Lee, Jaejin Cho, Kyung Chul Choi

School of Medicine

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

Canine mesenchymal stem cells (cMSCs) are gaining popularity in the veterinary field as a regenerative therapy. But, their limited culture lifespan makes it an obstacle for preclinical investigation and therapeutic use. In this study, primary canine adipose tissue-derived MSCs (PCAT-MSCs) were isolated from adipose tissue and were transfected with the SV40-T retrovirus resulting in a life-extended immortalized canine adipose tissue-derived MSCs (ICAT-MSCs). A comparison was made through the characterization of both PCAT-MSCs and ICAT-MSCs. Both showed a fibroblastic morphology; ICAT-MSCs showed a higher potential of colony formation compared with PCAT-MSCs and a reduced population doubling time; stem cell markers SOX2 and NANOG were expressed in both cell lines; karyotyping analysis showed no abnormalities in both PCAT-MSCs and ICAT-MSCs; both cell lines were CD90⁺, CD44 ⁺, and CD45 ⁻; both generated chondrogenic pellet; in osteogenic differentiation both showed upregulation of Osterix, a master transcriptome of osteogenesis, but in PCAT-MSCs, an upregulation of SOX2 was also observed. In conclusion, ICAT-MSCs showed similar characteristics with PCAT-MSCs, thus established as an easy to access platform for studies on better understanding about cMSCs nature.

Original language	English (US)
Pages (from-to)	16630-16642
Number of pages	13
Journal	Journal of Cellular Physiology
Volume	234
Issue number	9
DOIs	https://doi.org/10.1002/jcp.28338
State	Published - Sep 2019

Keywords

SV40-T
canine stem cells
differentiation
mesenchymal stem cells
osteogenesis

ASJC Scopus subject areas

Physiology
Clinical Biochemistry
Cell Biology

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10.1002/jcp.28338

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title = "Characterization of canine adipose tissue-derived mesenchymal stem cells immortalized by SV40-T retrovirus for therapeutic use",

abstract = "Canine mesenchymal stem cells (cMSCs) are gaining popularity in the veterinary field as a regenerative therapy. But, their limited culture lifespan makes it an obstacle for preclinical investigation and therapeutic use. In this study, primary canine adipose tissue-derived MSCs (PCAT-MSCs) were isolated from adipose tissue and were transfected with the SV40-T retrovirus resulting in a life-extended immortalized canine adipose tissue-derived MSCs (ICAT-MSCs). A comparison was made through the characterization of both PCAT-MSCs and ICAT-MSCs. Both showed a fibroblastic morphology; ICAT-MSCs showed a higher potential of colony formation compared with PCAT-MSCs and a reduced population doubling time; stem cell markers SOX2 and NANOG were expressed in both cell lines; karyotyping analysis showed no abnormalities in both PCAT-MSCs and ICAT-MSCs; both cell lines were CD90+, CD44 +, and CD45 −; both generated chondrogenic pellet; in osteogenic differentiation both showed upregulation of Osterix, a master transcriptome of osteogenesis, but in PCAT-MSCs, an upregulation of SOX2 was also observed. In conclusion, ICAT-MSCs showed similar characteristics with PCAT-MSCs, thus established as an easy to access platform for studies on better understanding about cMSCs nature.",

keywords = "SV40-T, canine stem cells, differentiation, mesenchymal stem cells, osteogenesis",

author = "Ayala-Cuellar, {Ana Patricia} and Kim, {Cho Won} and Hwang, {Kyung A.} and Kang, {Ji Houn} and Gabsang Lee and Jaejin Cho and Choi, {Kyung Chul}",

note = "Funding Information: This study was supported by the Global Research and Development Center (GRDC) Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT (2017K1A4A3014959). In addition, this study was also supported by a Grant (No. 317021–03‐1‐CG000) from Korea Institute of Planning and Evaluation for technology in Food, Agriculture and Forestry, Republic of Korea. Publisher Copyright: {\textcopyright} 2019 Wiley Periodicals, Inc.",

year = "2019",

month = sep,

doi = "10.1002/jcp.28338",

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AU - Ayala-Cuellar, Ana Patricia

AU - Kim, Cho Won

AU - Hwang, Kyung A.

AU - Kang, Ji Houn

AU - Lee, Gabsang

AU - Cho, Jaejin

AU - Choi, Kyung Chul

N1 - Funding Information: This study was supported by the Global Research and Development Center (GRDC) Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT (2017K1A4A3014959). In addition, this study was also supported by a Grant (No. 317021–03‐1‐CG000) from Korea Institute of Planning and Evaluation for technology in Food, Agriculture and Forestry, Republic of Korea. Publisher Copyright: © 2019 Wiley Periodicals, Inc.

PY - 2019/9

Y1 - 2019/9

N2 - Canine mesenchymal stem cells (cMSCs) are gaining popularity in the veterinary field as a regenerative therapy. But, their limited culture lifespan makes it an obstacle for preclinical investigation and therapeutic use. In this study, primary canine adipose tissue-derived MSCs (PCAT-MSCs) were isolated from adipose tissue and were transfected with the SV40-T retrovirus resulting in a life-extended immortalized canine adipose tissue-derived MSCs (ICAT-MSCs). A comparison was made through the characterization of both PCAT-MSCs and ICAT-MSCs. Both showed a fibroblastic morphology; ICAT-MSCs showed a higher potential of colony formation compared with PCAT-MSCs and a reduced population doubling time; stem cell markers SOX2 and NANOG were expressed in both cell lines; karyotyping analysis showed no abnormalities in both PCAT-MSCs and ICAT-MSCs; both cell lines were CD90+, CD44 +, and CD45 −; both generated chondrogenic pellet; in osteogenic differentiation both showed upregulation of Osterix, a master transcriptome of osteogenesis, but in PCAT-MSCs, an upregulation of SOX2 was also observed. In conclusion, ICAT-MSCs showed similar characteristics with PCAT-MSCs, thus established as an easy to access platform for studies on better understanding about cMSCs nature.

AB - Canine mesenchymal stem cells (cMSCs) are gaining popularity in the veterinary field as a regenerative therapy. But, their limited culture lifespan makes it an obstacle for preclinical investigation and therapeutic use. In this study, primary canine adipose tissue-derived MSCs (PCAT-MSCs) were isolated from adipose tissue and were transfected with the SV40-T retrovirus resulting in a life-extended immortalized canine adipose tissue-derived MSCs (ICAT-MSCs). A comparison was made through the characterization of both PCAT-MSCs and ICAT-MSCs. Both showed a fibroblastic morphology; ICAT-MSCs showed a higher potential of colony formation compared with PCAT-MSCs and a reduced population doubling time; stem cell markers SOX2 and NANOG were expressed in both cell lines; karyotyping analysis showed no abnormalities in both PCAT-MSCs and ICAT-MSCs; both cell lines were CD90+, CD44 +, and CD45 −; both generated chondrogenic pellet; in osteogenic differentiation both showed upregulation of Osterix, a master transcriptome of osteogenesis, but in PCAT-MSCs, an upregulation of SOX2 was also observed. In conclusion, ICAT-MSCs showed similar characteristics with PCAT-MSCs, thus established as an easy to access platform for studies on better understanding about cMSCs nature.

KW - SV40-T

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ER -

Characterization of canine adipose tissue-derived mesenchymal stem cells immortalized by SV40-T retrovirus for therapeutic use

Abstract

Keywords

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