Secreted adiponectin as a marker to evaluate in vitro the adipogenic differentiation of human mesenchymal stromal cells

Martella, Elisa; Bellotti, Chiara; Dozza, Barbara; Perrone, Sharon; Donati, Davide María; Lucarelli, Enrico

doi:10.1016/j.jcyt.2014.05.005

Cited by 38 publications

(26 citation statements)

References 41 publications

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“…In order to monitor the effects of differentiation in these experiments, we examined ADIPQ mRNA expression and correlated that to in vitro BAT activity. ADIPQ mRNA did not show a correlation with in vitro BAT activity ( Figure 2F), indicating that in vitro BAT activity was not dependent on the differentiation state of the adipocytes because ADIPQ is an important indicator of adipocyte differentiation (20,21).…”

Section: Zic1 Messenger Rna Negatively Correlates With In Vitro Bat Amentioning

confidence: 95%

Genetic Markers of Brown Adipose Tissue Identity and In Vitro Brown Adipose Tissue Activity in Humans

Nascimento

Sparks

Divoux

et al. 2017

Obesity

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Objective: Human brown adipose tissue (BAT) activity decreases with age and obesity. In addition to uncoupling protein 1 (UCP1), several genetic markers of BAT in humans have been published. However, the link between human BAT activity and genetic markers has been inadequately explored. Methods: White adipose tissue (WAT) and BAT biopsies were obtained from 16 patients undergoing deep neck surgery. In vitro differentiated adipocytes were used to measure norepinephrine-stimulated mitochondrial uncoupling as a measure of in vitro BAT activity. Gene expression was determined in adipose tissue biopsies. Results: Norepinephrine increased in vitro BAT activity in adipocytes derived from human BAT, and this increase was abolished by propranolol. Furthermore, in vitro BAT activity showed a negative correlation to age and BMI. UCP1 messenger RNA (mRNA) expression showed a positive correlation to in vitro BAT activity, while zinc finger protein of cerebellum 1 (ZIC1) mRNA showed a negative correlation to in vitro BAT activity. In human BAT biopsies, UCP1 mRNA showed negative correlations to age and BMI, while ZIC1 mRNA showed positive correlations to age and BMI. Conclusions: Differentiated adipocytes derived from human BAT maintain intrinsic characteristics of the donor. High ZIC1 mRNA does not necessarily reflect high BAT activity.

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Section: Zic1 Messenger Rna Negatively Correlates With In Vitro Bat Amentioning

confidence: 95%

Genetic Markers of Brown Adipose Tissue Identity and In Vitro Brown Adipose Tissue Activity in Humans

Nascimento

Sparks

Divoux

et al. 2017

Obesity

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“…Osteogenically and adipogenically induced aspirates were processed for quantitative estimation of the alkaline phosphatase (ALP) activities and Oil Red O staining, respectively. [40][41][42] Determination of ALP activity was performed by mixing 100 mL of osteogenically differentiated aspirate with an equal volume of substrate buffer (4 mg/mL 4-nitrophenyl phosphate-pNPP-mixed 1:1 with 4.8% 2-amino-2-methyl-1-propanol-2-AMP) and measuring OD 530nm as previously described. 40 Estimation of Oil Red O staining was performed by mixing 100 mL of adipogenically induced aspirate with 150 mL of staining solution (three volumes of Oil Red O 0.3% in 2-propanol and two volumes H 2 O) for 15 min at room temperature followed by dissolution in 100% 2-propanol and by measuring OD 530nm .…”

Section: Biochemical Assaysmentioning

confidence: 99%

Chondrogenic Differentiation Processes in Human Bone Marrow Aspirates upon rAAV-Mediated Gene Transfer and Overexpression of the Insulin-Like Growth Factor I

Frisch

Rey‐Rico

Venkatesan

et al. 2015

Tissue Engineering Part A

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Direct therapeutic gene transfer in marrow concentrates is an attractive strategy to conveniently enhance the chondrogenic differentiation processes as a means to improve the healing response of damaged articular cartilage upon reimplantation in sites of injury. In the present study, we evaluated the ability of the clinically adapted recombinant adeno-associated virus (rAAV) vectors to mediate overexpression of the insulin-like growth factor I (IGF-I) in human bone marrow aspirates that may modulate the proliferative, anabolic activities, and chondrogenic differentiation potential in such samples in vitro. The results demonstrate that successful, significant rAAV-mediated IGF-I gene transfer and expression were achieved in transduced aspirates (up to 105.9 -35.1 pg rhIGF-I/mg total proteins) over time (21 days) at very high levels (*80% of cells expressing the candidate IGF-I transgene), leading to increased levels of proliferation, matrix synthesis, and chondrogenic differentiation over time compared with the control (lacZ) condition. Treatment with the candidate IGF-I vector also stimulated the hypertrophic and osteogenic differentiation processes in the aspirates, suggesting that the regulation of IGF-I expression through rAAV will be a prerequisite for future translation of the approach in vivo. However, these findings show the possible benefits of this vector class to directly modify marrow concentrates as a convenient tool for strategies that aim at improving the repair of articular cartilage lesions.

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“…Food intake for the four groups over the 14 days of treatment was similar (Table 1). To evaluate cell turnover, we measured the mRNA levels of Pref1 (pre-adipocyte marker) because its expression is linked to adipogenesis inhibition (O'Connell et al 2011) and the mRNA levels of Adiponectin, which have been used as a marker of mature adipocytes found in the SVF (Martella et al 2014). There was no change in Pref1 expression in the SVF from RPAT samples at days 7 and 14 ( Fig.…”

Section: Resultsmentioning

confidence: 96%

Cancer cachexia differentially regulates visceral adipose tissue turnover

Franco

Lopes

Henriques

et al. 2017

Journal of Endocrinology

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Cancer cachexia (CC) is a progressive metabolic syndrome that is marked by severe body weight loss. Metabolic disarrangement of fat tissues is a very early event in CC, followed by adipose tissue (AT) atrophy and remodelling. However, there is little information regarding the possible involvement of cellular turnover in this process. Thus, in this study, we evaluated the effect of CC on AT turnover and fibrosis of mesenteric (MEAT) and retroperitoneal (RPAT) adipose tissue depots as possible factors that contribute to AT atrophy. CC was induced by a subcutaneous injection of Walker tumour cells (2 × 10) in Wistar rats, and control animals received only saline. The experimental rats were randomly divided into four experimental groups: 0 days, 4 days, 7 days and 14 days after injection. AT turnover was analysed according to the Pref1/Adiponectin ratio of gene expression from the stromal vascular fraction and pro-apoptotic CASPASE3 and CASPASE9 from MEAT and RPAT. Fibrosis was verified according to the total collagen levels and expression of extracellular matrix genes. AT turnover was verified by measurements of lipolytic protein expression. We found that the Pref1/Adiponectin ratio was decreased in RPAT (81.85%, P < 0.05) with no changes in MEAT compared with the respective controls. CASPASE3 and CASPASE9 were activated on day 14 only in RPAT. Collagen was increased on day 7 in RPAT (127%) and MEAT (4.3-fold). The Collagen1A1, Collagen3A1, Mmp2 and Mmp9 mRNA levels were upregulated only in MEAT in CC. Lipid turnover was verified in RPAT and was not modified in CC. We concluded that the results suggest that CC affects RPAT cellular turnover, which may be determinant for RPAT atrophy.

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Secreted adiponectin as a marker to evaluate in vitro the adipogenic differentiation of human mesenchymal stromal cells

Cited by 38 publications

References 41 publications

Genetic Markers of Brown Adipose Tissue Identity and In Vitro Brown Adipose Tissue Activity in Humans

Genetic Markers of Brown Adipose Tissue Identity and In Vitro Brown Adipose Tissue Activity in Humans

Chondrogenic Differentiation Processes in Human Bone Marrow Aspirates upon rAAV-Mediated Gene Transfer and Overexpression of the Insulin-Like Growth Factor I

Cancer cachexia differentially regulates visceral adipose tissue turnover

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