Characterization of immortalized human brown and white pre-adipocyte cell models from a single donor

Lasse K Markussen, Marie S Isidor, Peter Breining, Elise S Andersen, Nanna E Rasmussen, Louise I Petersen, Steen B Pedersen, Bjørn Richelsen, Jacob B Hansen

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Resumé

Brown adipose tissue with its constituent brown adipocytes is a promising therapeutic target in metabolic disorders due to its ability to dissipate energy and improve systemic insulin sensitivity and glucose homeostasis. The molecular control of brown adipocyte differentiation and function has been extensively studied in mice, but relatively little is known about such regulatory mechanisms in humans, which in part is due to lack of human brown adipose tissue derived cell models. Here, we used retrovirus-mediated overexpression to stably integrate human telomerase reverse transcriptase (TERT) into stromal-vascular cell fractions from deep and superficial human neck adipose tissue biopsies from the same donor. The brown and white pre-adipocyte cell models (TERT-hBA and TERT-hWA, respectively) displayed a stable proliferation rate and differentiation until at least passage 20. Mature TERT-hBA adipocytes expressed higher levels of thermogenic marker genes and displayed a higher maximal respiratory capacity than mature TERT-hWA adipocytes. TERT-hBA adipocytes were UCP1-positive and responded to β-adrenergic stimulation by activating the PKA-MKK3/6-p38 MAPK signaling module and increasing thermogenic gene expression and oxygen consumption. Mature TERT-hWA adipocytes underwent efficient rosiglitazone-induced 'browning', as demonstrated by strongly increased expression of UCP1 and other brown adipocyte-enriched genes. In summary, the TERT-hBA and TERT-hWA cell models represent useful tools to obtain a better understanding of the molecular control of human brown and white adipocyte differentiation and function as well as of browning of human white adipocytes.

OriginalsprogEngelsk
TidsskriftPLOS ONE
Vol/bind12
Udgave nummer9
Sider (fra-til)e0185624
ISSN1932-6203
DOI
StatusUdgivet - sep. 2017
Udgivet eksterntJa

Fingeraftryk

White Adipocytes
Telomerase
telomerase
RNA-directed DNA polymerase
adipocytes
Brown Adipocytes
Adipocytes
cells
rosiglitazone
Tissue
brown adipose tissue
Genes
Retroviridae
Biopsy
p38 Mitogen-Activated Protein Kinases
Stromal Cells
Gene expression
Oxygen Consumption
Adrenergic Agents
Insulin Resistance

Citer dette

Markussen, L. K., Isidor, M. S., Breining, P., Andersen, E. S., Rasmussen, N. E., Petersen, L. I., ... Hansen, J. B. (2017). Characterization of immortalized human brown and white pre-adipocyte cell models from a single donor. PLOS ONE, 12(9), e0185624. https://doi.org/10.1371/journal.pone.0185624
Markussen, Lasse K ; Isidor, Marie S ; Breining, Peter ; Andersen, Elise S ; Rasmussen, Nanna E ; Petersen, Louise I ; Pedersen, Steen B ; Richelsen, Bjørn ; Hansen, Jacob B. / Characterization of immortalized human brown and white pre-adipocyte cell models from a single donor. I: PLOS ONE. 2017 ; Bind 12, Nr. 9. s. e0185624.
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title = "Characterization of immortalized human brown and white pre-adipocyte cell models from a single donor",
abstract = "Brown adipose tissue with its constituent brown adipocytes is a promising therapeutic target in metabolic disorders due to its ability to dissipate energy and improve systemic insulin sensitivity and glucose homeostasis. The molecular control of brown adipocyte differentiation and function has been extensively studied in mice, but relatively little is known about such regulatory mechanisms in humans, which in part is due to lack of human brown adipose tissue derived cell models. Here, we used retrovirus-mediated overexpression to stably integrate human telomerase reverse transcriptase (TERT) into stromal-vascular cell fractions from deep and superficial human neck adipose tissue biopsies from the same donor. The brown and white pre-adipocyte cell models (TERT-hBA and TERT-hWA, respectively) displayed a stable proliferation rate and differentiation until at least passage 20. Mature TERT-hBA adipocytes expressed higher levels of thermogenic marker genes and displayed a higher maximal respiratory capacity than mature TERT-hWA adipocytes. TERT-hBA adipocytes were UCP1-positive and responded to β-adrenergic stimulation by activating the PKA-MKK3/6-p38 MAPK signaling module and increasing thermogenic gene expression and oxygen consumption. Mature TERT-hWA adipocytes underwent efficient rosiglitazone-induced 'browning', as demonstrated by strongly increased expression of UCP1 and other brown adipocyte-enriched genes. In summary, the TERT-hBA and TERT-hWA cell models represent useful tools to obtain a better understanding of the molecular control of human brown and white adipocyte differentiation and function as well as of browning of human white adipocytes.",
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Markussen, LK, Isidor, MS, Breining, P, Andersen, ES, Rasmussen, NE, Petersen, LI, Pedersen, SB, Richelsen, B & Hansen, JB 2017, 'Characterization of immortalized human brown and white pre-adipocyte cell models from a single donor', PLOS ONE, bind 12, nr. 9, s. e0185624. https://doi.org/10.1371/journal.pone.0185624

Characterization of immortalized human brown and white pre-adipocyte cell models from a single donor. / Markussen, Lasse K; Isidor, Marie S; Breining, Peter; Andersen, Elise S; Rasmussen, Nanna E; Petersen, Louise I; Pedersen, Steen B; Richelsen, Bjørn; Hansen, Jacob B.

I: PLOS ONE, Bind 12, Nr. 9, 09.2017, s. e0185624.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Characterization of immortalized human brown and white pre-adipocyte cell models from a single donor

AU - Markussen, Lasse K

AU - Isidor, Marie S

AU - Breining, Peter

AU - Andersen, Elise S

AU - Rasmussen, Nanna E

AU - Petersen, Louise I

AU - Pedersen, Steen B

AU - Richelsen, Bjørn

AU - Hansen, Jacob B

PY - 2017/9

Y1 - 2017/9

N2 - Brown adipose tissue with its constituent brown adipocytes is a promising therapeutic target in metabolic disorders due to its ability to dissipate energy and improve systemic insulin sensitivity and glucose homeostasis. The molecular control of brown adipocyte differentiation and function has been extensively studied in mice, but relatively little is known about such regulatory mechanisms in humans, which in part is due to lack of human brown adipose tissue derived cell models. Here, we used retrovirus-mediated overexpression to stably integrate human telomerase reverse transcriptase (TERT) into stromal-vascular cell fractions from deep and superficial human neck adipose tissue biopsies from the same donor. The brown and white pre-adipocyte cell models (TERT-hBA and TERT-hWA, respectively) displayed a stable proliferation rate and differentiation until at least passage 20. Mature TERT-hBA adipocytes expressed higher levels of thermogenic marker genes and displayed a higher maximal respiratory capacity than mature TERT-hWA adipocytes. TERT-hBA adipocytes were UCP1-positive and responded to β-adrenergic stimulation by activating the PKA-MKK3/6-p38 MAPK signaling module and increasing thermogenic gene expression and oxygen consumption. Mature TERT-hWA adipocytes underwent efficient rosiglitazone-induced 'browning', as demonstrated by strongly increased expression of UCP1 and other brown adipocyte-enriched genes. In summary, the TERT-hBA and TERT-hWA cell models represent useful tools to obtain a better understanding of the molecular control of human brown and white adipocyte differentiation and function as well as of browning of human white adipocytes.

AB - Brown adipose tissue with its constituent brown adipocytes is a promising therapeutic target in metabolic disorders due to its ability to dissipate energy and improve systemic insulin sensitivity and glucose homeostasis. The molecular control of brown adipocyte differentiation and function has been extensively studied in mice, but relatively little is known about such regulatory mechanisms in humans, which in part is due to lack of human brown adipose tissue derived cell models. Here, we used retrovirus-mediated overexpression to stably integrate human telomerase reverse transcriptase (TERT) into stromal-vascular cell fractions from deep and superficial human neck adipose tissue biopsies from the same donor. The brown and white pre-adipocyte cell models (TERT-hBA and TERT-hWA, respectively) displayed a stable proliferation rate and differentiation until at least passage 20. Mature TERT-hBA adipocytes expressed higher levels of thermogenic marker genes and displayed a higher maximal respiratory capacity than mature TERT-hWA adipocytes. TERT-hBA adipocytes were UCP1-positive and responded to β-adrenergic stimulation by activating the PKA-MKK3/6-p38 MAPK signaling module and increasing thermogenic gene expression and oxygen consumption. Mature TERT-hWA adipocytes underwent efficient rosiglitazone-induced 'browning', as demonstrated by strongly increased expression of UCP1 and other brown adipocyte-enriched genes. In summary, the TERT-hBA and TERT-hWA cell models represent useful tools to obtain a better understanding of the molecular control of human brown and white adipocyte differentiation and function as well as of browning of human white adipocytes.

KW - Adipocytes/cytology

KW - Adipose Tissue, Brown/cytology

KW - Adipose Tissue, White/cytology

KW - Biopsy

KW - Cell Line, Transformed

KW - Colforsin/pharmacology

KW - Humans

KW - Isoproterenol/pharmacology

KW - Neck

KW - Retroviridae/genetics

KW - Telomerase/genetics

KW - Thermogenesis

KW - Thiazolidinediones/pharmacology

KW - Tissue Donors

U2 - 10.1371/journal.pone.0185624

DO - 10.1371/journal.pone.0185624

M3 - Journal article

VL - 12

SP - e0185624

JO - P L o S One

JF - P L o S One

SN - 1932-6203

IS - 9

ER -