Synergy Between Choroid Plexus Epithelial Cell-Conditioned Medium and Knockout Serum Replacement Converts Human Adipose-Derived Stem Cells to Dopamine-Secreting Neurons

Abstract

Human adipose-derived stem cells (hADSCs) have great capacity to differentiate into mesodermal origins as well as nonmesodermal lineages, including neural cells. This valuable feature paves the way for the therapeutic application of hADSCs for neurodegenerative maladies such as Parkinson's disease (PD). We tested the capacity of choroid plexus epithelial cell-conditioned medium (CPEC-CM) alone or cocktailed with knockout serum (KS) to induce dopaminergic (DAergic) differentiation of hADSCs. To this end, hADSCs from lipoaspirate were phenotypically characterized and shown to maintain mesodermal multipotency so that selected media easily differentiated them into osteoblasts, chondrocytes, and adipocytes. To begin inducing hADSC neuronal differentiation, we isolated CPECs from rat brain and expanded them in culture to obtain CPEC-CM. We then treated hADSCs with optimized quantities of collected CPEC-CM, KS, or both. The ADSCs treated with either CPEC-CM or CPEC-CM and KS displayed morphological changes typical of neuron-like phenotypes. As revealed by reverse transcription polymerase chain reaction (RT-PCR), quantitative real-time PCR (qPCR), and immunostaining analyses, hADSCs cotreated with CPEC-CM and KS expressed significantly higher levels of neuronal and DAergic markers in comparison with single-treated groups. Moreover, the hADSCs began expressing dopamine-biosynthesizing enzymes mainly after cotreatment with CPEC-CM and KS. Consequently, only cotreated hADSCs were capable of synthesizing and releasing dopamine detectable by high-performance liquid chromatography (HPLC). Finally, hADSCs growing in an ordinary medium were found positive for astrocytic marker glial fibrillary acidic protein (GFAP), but stopped GFAP expression on either single or cotreatments. These combined results suggest that CPEC-CM and KS can synergize to remarkably augment DAergic induction of hADSCs, an effect that has implications for cell replacement therapy for PD and related disorders.

Functional dopaminergic neurons derived from human chorionic mesenchymal stem cells ameliorate striatal atrophy and improve behavioral deficits in Parkinsonian rat model

“…Our findings confirmed that implantation of HCMSCs‐derived DA neuron‐like cells into the striatum of a rat model of PD could noticeably impede morphological and behavioral alterations, following 6‐OHDA‐induced lesion. In addition, our two recent experiments displayed DA differentiation of adipose‐derived MSCs under conditioned‐medium treatment (Boroujeni, Gardaneh, Shahriari, Aliaghaei, & Hasani, ; Rad et al, ). Moreover, Bjorklund et al (Björklund et al, ) disclosed that ES cells could develop into functional DA neuron‐like cells, leading to decline in motor asymmetries, accompanied by normalized DA presynaptic markers in damaged animal brain, following cell transplant.…”

Differentiation of human olfactory system‐derived stem cells into dopaminergic neuron‐like cells: A comparison between olfactory bulb and mucosa as two sources of stem cells

“…Cell transplantation is believed to be a promising therapeutic approach in the treatment process of neurodegenerative disorders such as PD when there are no definite pharmaceutical therapies . However, tumorigenesis and ethical issues must always be considered when it comes to cell transplantation …”

“…[28][29][30] The current study aimed to make a comparison between the differentiation capacity of the OE-MSCs and OB-NSCs into DA-ergic neuron-like cells under the inductive effect of the transforming growth factor β (TGF-β) differentiation factor, which is substantially needed to induce DA-ergic cells in the mesencephalon and indeed promotes the DA-ergic neurons' survival. 47…”

Reprogramming by Cytosolic Extract of Human Embryonic Stem Cells to Improve Dopaminergic Differentiation Potential of Human Adipose Tissue-derived Stem Cells

“…Han et al, 2014;Safford et al, 2002). In recent years, several studies have documented dopaminergic differentiation of hADSCs (Boroujeni, Gardaneh, Shahriari, Aliaghaei, & Hasani, 2017;Faghih, Javeri, Amini, & Taha, 2019;Marei et al, 2018;Singh et al, 2017;Soheilifar et al, 2018) which raises the hope of using these cells for the treatment of Parkinson's disease. In a recent work, we successfully induced differentiation of hADSCs to dopaminergic neurons by a growth factor-cocktail consisting of SHH, bFGF, FGF8 and BDNF.…”