Isolation of adipose tissue mesenchymal stem cells without tissue destruction: A non-enzymatic method

Ghorbani, Ahmad; Jalali, Seyed Amir; Varedi, Masoumeh

doi:10.1016/j.tice.2013.11.002

Cited by 63 publications

(47 citation statements)

References 17 publications

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“…Subcutaneous inguinal fat deposits were resected and under laminar hood, the fat tissue was minced into 1–2 mm pieces by means of a sterile scalpel [17]. The tissue pieces were incubated at 37°C for 60 minutes in PBS containing 2 mg/ml collagenase meanwhile being shaken (60 cycles/min) [18,19]. After centrifugation (300 g for 8 minutes), the floated lipid layer was discarded and the stroma-vascular fraction was collected, washed and re-suspended in DMEM medium supplemented with 10% FBS, 100 units/ml penicillin and 100 μg/ml streptomycin [19,20].…”

Section: Methodsmentioning

confidence: 99%

“…The tissue pieces were incubated at 37°C for 60 minutes in PBS containing 2 mg/ml collagenase meanwhile being shaken (60 cycles/min) [18,19]. After centrifugation (300 g for 8 minutes), the floated lipid layer was discarded and the stroma-vascular fraction was collected, washed and re-suspended in DMEM medium supplemented with 10% FBS, 100 units/ml penicillin and 100 μg/ml streptomycin [19,20]. The cells were seeded into tissue culture flask and passaged when 60–80% confluent and used at passages three to six.…”

Section: Methodsmentioning

confidence: 99%

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The effect of adipose derived stromal cells on oxidative stress level, lung emphysema and white blood cells of guinea pigs model of chronic obstructive pulmonary disease

et al. 2014

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BackgroundChronic obstructive pulmonary disease (COPD) is a worldwide epidemic disease and a major cause of death and disability. The present study aimed to elucidate pharmacological effects of adipose derived stromal cells (ASCs) on pathological and biochemical factors in a guinea pig model of COPD. Guinea pigs were randomized into 5 groups including: Control, COPD, COPD + intratracheal delivery of PBS as a vehicle (COPD-PBS), COPD + intratracheal delivery of ASCs (COPD-ITASC) and COPD + intravenous injection of ASCs (COPD-IVASC). COPD was induced by exposing animals to cigarette smoke for 3 months. Cell therapy was performed immediately after the end of animal exposure to cigarette smoke and 14 days after that, white blood cells, oxidative stress indices and pathological changes of the lung were measured.ResultsCompared with control group, emphysema was clearly observed in the COPD and COPD-PBS groups (p < 0.001). Lung histopathologic changes of COPD-ITASC and COPD-IVASC groups showed non-significant improvement compared to COPD-PBS group. The COPD-ITASC group showed a significant increase in total WBC compared to COPD-PBS group but there was not a significant increase in this regard in COPD-IVASC group. The differential WBC showed no significant change in number of different types of leukocytes. The serum level of malondialdehyde (MDA) significantly decreased but thiol groups of broncho-alveolar lavage fluid (BALF) increased in both cell treated groups (p < 0.05 for all cases). Weight of animals decreased during smoke exposure and improved after PBS or cell therapy. However, no significant change was observed between the groups receiving PBS and the ones receiving ASCs.ConclusionCell therapy with ASCs can help in reducing oxidative damage during smoking which may collectively hold promise in attenuation of the severity of COPD although the lung structural changes couldn’t be ameliorated with these pharmacological therapeutic methods.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

The effect of adipose derived stromal cells on oxidative stress level, lung emphysema and white blood cells of guinea pigs model of chronic obstructive pulmonary disease

et al. 2014

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“…Notably, this study reported no major differences in cell characteristics isolated from enzymatic and non-enzymatic methods; however, cellular yield was higher in the enzymatically digested method. In another recent study, the mesenchymal stem cells (MSCs) obtained from harvested adipose tissue of animal or human were pluripotent and successfully differentiated into adipocyte and osteoblasts [106]. Various commercial mechanical devices have been developed to process adipose tissue; which uses forces, such as pressure, centrifugal force, shear force, radiation, and ultrasound, etc.…”

Section: Adipose Tissues As Source Of Ascsmentioning

confidence: 99%

Revisiting the Advances in Isolation, Characterization and Secretome of Adipose-Derived Stromal/Stem Cells

Dubey

Mishra²,

Dubey

et al. 2018

IJMS

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Adipose-derived stromal/stem cells (ASCs) seems to be a promising regenerative therapeutic agent due to the minimally invasive approach of their harvest and multi-lineage differentiation potential. The harvested adipose tissues are further digested to extract stromal vascular fraction (SVF), which is cultured, and the anchorage-dependent cells are isolated in order to characterize their stemness, surface markers, and multi-differentiation potential. The differentiation potential of ASCs is directed through manipulating culture medium composition with an introduction of growth factors to obtain the desired cell type. ASCs have been widely studied for its regenerative therapeutic solution to neurologic, skin, wound, muscle, bone, and other disorders. These therapeutic outcomes of ASCs are achieved possibly via autocrine and paracrine effects of their secretome comprising of cytokines, extracellular proteins and RNAs. Therefore, secretome-derivatives might offer huge advantages over cells through their synthesis and storage for long-term use. When considering the therapeutic significance and future prospects of ASCs, this review summarizes the recent developments made in harvesting, isolation, and characterization. Furthermore, this article also provides a deeper insight into secretome of ASCs mediating regenerative efficacy.

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“…Briefly, the cells were washed twice with PBS and fixed with 10% formalin for 30 min. After an additional washing with PBS, the cells were incubated with Oil Red O solution for 20 min [21,23]. Excess stain was removed by washing with distilled water and the stained cells were photographed.…”

Section: Methodsmentioning

confidence: 99%

Metabolic effects of newly synthesized phosphodiesterase-3 inhibitor 6-[4-(4-methylpiperidin-1-yl)-4-oxobutoxy]-4-methylquinolin-2(1H)-one on rat adipocytes

et al. 2015

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BackgroundClinical use of selective PDE3 inhibitors as cardiotonic agents is limited because of their chronotropic and lipolytic side effects. In our previous work, we synthesized a new PDE3 inhibitor named MC2 (6-[4-(4-methylpiperidin-1-yl)-4-oxobutoxy]-4-methylquinolin-2(1H)-one) which produced a high positive inotropic action with a negative chronotropic effect. This work was done to evaluate the effects of MC2 on adipocytes and compare its effects with those of amrinone and cilostamide.MethodsPreadipocytes were isolated from rat adipose tissue and differentiated to adipocyte in the presence of cilostamide, amrinone or MC2. Lipolysis and adipogenesis was evaluated by measuring glycerol level and Oil Red O staining, respectively. Adipocyte proliferation and apoptosis were determined with MTT assay and Annexin V/PI staining, respectively.ResultsDifferentiation to adipocyte was induced by amrinone but not by cilostamide or MC2. Basal and isoproterenol-stimulated lipolysis significantly increased by cilostamide (p < 0.05). Similarly, amrinone enhanced the stimulated lipolysis (p < 0.01). On the other hand, MC2 significantly decreased both adipogenesis (p < 0.05) and stimulated lipolysis (p < 0.001). Also, incubation of differentiated adipocytes with MC2 caused the loss of cell viability, which was associated with the elevation in apoptotic rate (p < 0.05).ConclusionOur data indicate that selective PDE3 inhibitors produce differential effects on adipogenesis and lipolysis. MC2 has proapoptotic and antilipolytic effects on adipocytes and does not stimulate adipogenesis. Therefore, in comparison with the clinically available selective PDE3 inhibitors, MC2 has lowest metabolic side effects and might be a good candidate for treatment of congestive heart failure.

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Isolation of adipose tissue mesenchymal stem cells without tissue destruction: A non-enzymatic method

Cited by 63 publications

References 17 publications

The effect of adipose derived stromal cells on oxidative stress level, lung emphysema and white blood cells of guinea pigs model of chronic obstructive pulmonary disease

The effect of adipose derived stromal cells on oxidative stress level, lung emphysema and white blood cells of guinea pigs model of chronic obstructive pulmonary disease

Revisiting the Advances in Isolation, Characterization and Secretome of Adipose-Derived Stromal/Stem Cells

Metabolic effects of newly synthesized phosphodiesterase-3 inhibitor 6-[4-(4-methylpiperidin-1-yl)-4-oxobutoxy]-4-methylquinolin-2(1H)-one on rat adipocytes

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