Simplified culture techniques for growth and differentiation of murine and human pre-adipocytes for translational applications

Herrera-Herrera, Mireya L.; Zapata-Bustos, Rocio; Salazar‐Olivo, Luis A.

doi:10.1080/14653240802495963

Cited by 10 publications

(4 citation statements)

References 28 publications

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“…Totally, 36 articles were identified. Of those, 30 were included in the review: there were 12 case reports [1,9,10,11,26,27,28,29,30,31,32,33,34], 12 reports on basic science or animal studies [4,12,13,35,36,37,38,39,40,41,42,43,44], 2 case series [14,45] and 4 systematic reviews [13,38,46,47]. Totally, more than 60 patients were analysed; 25 articles were studied as a full-text version.…”

Section: Methodsmentioning

confidence: 99%

Adipose-Derived Stromal Cells from Lipomas: Isolation, Characterisation and Review of the Literature

Tremp

Menzi²,

Tchang³

et al. 2016

Pathobiology

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Objective: The aim of this study was to characterize adipose-derived stromal cells (ADSCs) from patients diagnosed with multiple symmetric lipomatosis (MSL) in order to obtain potentially new insights into the pathophysiology, pathogenesis and treatment of this disease. Methods: Cells from the stromal vascular fraction were analysed by the colony-forming efficiency assay and flow cytometry using standard markers. Moreover, the power of adipogenic plasticity was evaluated. Finally, a literature review was performed from 1982 to 2015 using the US National Institutes of Health's PubMed database. Results: Three European-descent patients diagnosed with either MSL type I or II could be identified for analysis. The resulting mean colony-forming efficiency assay was 14.3 ± 5%. Flow-cytometric analysis of the ADSCs revealed high levels of CD34 (70 ± 9%), CD45 (37 ± 13%) and CD73 (55.8 ± 14%), whereas low levels of CD31 (16.8 ± 14%) and CD105 (5.8 ± 0.7%) were detected. Furthermore, ADSCs showed a strong adipogenic potential, which is in line with the literature review. The stem cell pool in lipoma shows several alterations in biological activities, such as proliferation, apoptosis and stemness. Conclusions: ADSCs from lipoma may be interesting in the application of regenerative medicine. We discuss possible molecular treatment options to regulate their activities at the source of the MSL.

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Section: Methodsmentioning

confidence: 99%

Adipose-Derived Stromal Cells from Lipomas: Isolation, Characterisation and Review of the Literature

Tremp

Menzi²,

Tchang³

et al. 2016

Pathobiology

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“…Human preadipocytes were isolated by collagenase digestion (27) of sc adipose tissue samples obtained from nondiabetic patients undergoing elective cosmetic surgery, who gave their informed consent for the use of discarded tissues and with the approval of the corresponding Research Ethic Committee. DMEM, Leibovitz L15 (L15) medium and fetal bovine serum (FBS) were from GIBCO BRL (Grand Island, NY), whereas calf serum was from HyClone (Logan, UT).…”

Section: Reagents and Cell Linesmentioning

confidence: 99%

Isoorientin Reverts TNF-α-Induced Insulin Resistance in Adipocytes Activating the Insulin Signaling Pathway

et al. 2012

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Isoorientin (ISO) is a plant C-glycosylflavonoid with purported antidiabetic effects but unexplored mechanisms of action. To gain insight into its antidiabetic mechanisms, we assayed nontoxic ISO concentrations on the 2-(N-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl) amino)-2-deoxy-D-glucose (2-NBDG) uptake by murine 3T3-F442A and human sc adipocytes. In insulin-sensitive adipocytes, ISO stimulated the 2-NBDG uptake by 210% (murine) and 67% (human), compared with insulin treatment. Notably, ISO also induced 2-NBDG uptake in murine (139%) and human (60%) adipocytes made resistant to insulin by treatment with TNF-␣, compared with the incorporation induced in these cells by rosiglitazone. ISO induction of glucose uptake in adipocytes was abolished by inhibitors of the insulin signaling pathway. These inhibitors also blocked the proper phosphorylation of insulin signaling pathway components induced by ISO in both insulin-sensitive and insulinresistant adipocytes. Additionally, ISO stimulated the transcription of genes encoding components of insulin signaling pathway in murine insulin-sensitive and insulin-resistant adipocytes. In summary, we show here that ISO exerts its antidiabetic effects by activating the insulin signaling pathway in adipocytes, reverts the insulin resistance caused in these cells by TNF-␣ by stimulating the proper phosphorylation of proteins in this signaling pathway, and induces the expression of genes encoding these proteins. (Endocrinology 153: 5222-5230, 2012)

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“…Confluent cultures of 3T3-L1 preadipocytes were induced to adipose differentiation with adipogenic medium (AM; L15 medium added with 10% (v/v) fetal bovine serum (FBS), 0.25 µM dexamethasone, 0.1 mM 3-isobutyl-1-methylxanthine, 1 µM d-biotin, 80 U/ml penicillin, 80 µg/ml streptomycin and 5 µg/ml insulin) (Herrera-Herrera et al, 2009). After three days, the cells were fed with maintenance medium (MM; L15 added with 10% (v/v) FBS, 100 nM insulin, 1 µM d-biotin, 80 U/ml penicillin and 80 µg/ml streptomycin).…”

Section: Methodsmentioning

confidence: 99%

Expression of the Biologically Active Insulin Analog SCI-57 in Nicotiana Benthamiana

Muñoz-Talavera¹,

Gómez-Lim²,

Salazar‐Olivo³

et al. 2019

Front. Pharmacol.

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Diabetes mellitus is a growing problem worldwide; however, only 23% of low-income countries have access to insulin, and ironically it costs higher in such countries than high-income ones. Therefore, new strategies for insulin and insulin analogs production are urgently required to improve low-cost access to therapeutic products, so as to contain the diabetes epidemic. SCI-57 is an insulin analog with a greater affinity for the insulin receptor and lower thermal degradation than native insulin. It also shows native mitogenicity and insulin-like biological activity. In this work, SCI-57 was transiently expressed in the Nicotiana benthamiana (Nb) plant, and we also evaluated some of its relevant biological effects. An expression plasmid was engineered to translate an N-terminal ubiquitin and C-terminal endoplasmic reticulum-targeting signal KDEL, in order to increase protein expression and stability. Likewise, the effect of co-expression of influenza M2 ion channel (M2) on the expression of insulin analog SCI-57 (SCI-57/M2) was evaluated. Although using M2 increases yield, it tends to alter the SCI-57 amino acid sequence, possibly promoting the formation of oligomers. Purification of SCI-57 was achieved by FPLC cation exchange and ultrafiltration of N. benthamiana leaf extract (NLE). SCI-57 exerts its anti-diabetic properties by stimulating glucose uptake in adipocytes, without affecting the lipid accumulation process. Expression of the insulin analog in agroinfiltrated plants was confirmed by SDS-PAGE, RP-HPLC, and MS. Proteome changes related to the expression of heterologous proteins on N. benthamiana were not observed; up-regulated proteins were related to the agroinfiltration process. Our results demonstrate the potential for producing a biologically active insulin analog, SCI-57, by transient expression in Nb.

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Simplified culture techniques for growth and differentiation of murine and human pre-adipocytes for translational applications

Cited by 10 publications

References 28 publications

Adipose-Derived Stromal Cells from Lipomas: Isolation, Characterisation and Review of the Literature

Adipose-Derived Stromal Cells from Lipomas: Isolation, Characterisation and Review of the Literature

Isoorientin Reverts TNF-α-Induced Insulin Resistance in Adipocytes Activating the Insulin Signaling Pathway

Expression of the Biologically Active Insulin Analog SCI-57 in Nicotiana Benthamiana

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