Adipose-derived stem cells: An optimized protocol for isolation and proliferation

Ghiasi, Mahdieh; Qomi, Reza Tabatabaei; Kalhor, Neda; Sheykhhasan, Mohsen

doi:10.5530/ami.2016.1.25

Cited by 5 publications

(1 citation statement)

References 2 publications

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“…Then, we confirmed their stem cell identity by differentiating them to adipocytes, chondrocytes, neural cells, and osteocytes. The transduction of hADSCs with lentiviral vectors was high efficient with low toxicity (Ghiasi, Qomi, Kalhor, & Sheykhhasan, 2016; Morizono et al, 2003). We carried out this by the creation of lentiviral vectors through cotransfection of three plasmids in HEK‐293T and concentrate the assembled lentiviruses with PEG6000.…”

Section: Discussionmentioning

confidence: 99%

Impact of IFN‐β and LIF overexpression on human adipose‐derived stem cells properties

Ashja-Arvan

Dehbashi

Eslami

et al. 2020

Journal Cellular Physiology

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Adipose‐derived stem cells (ADSCs) are a subset of mesenchymal stem cells that their therapeutic effects in various diseases make them an interesting tool in cell therapy. In the current study, we aimed to overexpress interferon‐β (IFN‐β) and leukemia inhibitory factor (LIF) cytokines in human ADSCs to evaluate the impact of this overexpression on human ADSCs properties. Here, we designed a construct containing IFN‐β and LIF and then, transduced human adipose‐derived stem cells (hADSCs) by this construct via a lentiviral vector (PCDH‐513B). We assessed the ability of long‐term expression of the transgene in transduced cells by western blot analysis and enzyme‐linked immunosorbent assay techniques on Days 15, 45, and 75 after transduction. For the evaluation of stem cell properties, flow cytometry and differentiation assays were performed. Finally, the MTT assay was done to assess the proliferation of transduced cells compares to controls. Our results showed high‐efficiency transduction with highest expression rates on Day 75 after transduction which were 70 pg/ml for IFN‐β and 77.9 pg/ml for LIF in comparison with 25.60 pg/ml and 27.63 pg/ml, respectively, in untransduced cells (p = .0001). Also, transduced cells expressed a high level of ADSCs surface markers and successfully differentiated into adipocytes, chondrocytes, neural cells, and osteocytes besides the preservation rate of proliferation near untreated cells (p = .88). All in all, we successfully constructed an hADSC population stably overexpressed IFN‐β and LIF cytokines. Considering the IFN‐β and LIF anti‐inflammatory and neuroprotective effects as well as immune‐regulatory properties of hADSCs, the obtained cells of this study could be subjected for further evaluations in experimental autoimmune encephalomyelitis mice model.

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Section: Discussionmentioning

confidence: 99%