Effective &lt;i&gt;in vitro&lt;/i&gt; differentiation of adipose-derived stem cells into Schwann-like cells with folic acid supplementation

Chen, Shuhai; Ikemoto, Tetsuya; Tokunaga, Takuya; Okikawa, Shohei; Miyazaki, Katsuki; Tokuda, Kazunori; Yamada, Shinichiro; Saito, Yu; Imura, Satoru; Morine, Yuji; Shimada, Mitsuo

doi:10.2152/jmi.68.347

Cited by 6 publications

(7 citation statements)

References 42 publications

(52 reference statements)

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“…Human ADSCs were differentiated into SLCs using our protocol according to our previous report 11 . Representative images show the morphological changes in the 3D SLC group during the differentiation process ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…We differentiated ADSCs into 3D SLC spheroids using a modified protocol with an RCP μ-piece for PNI treatment. The enhanced protocol uses non-gene editing methods, and its advantages include its ability to induce ADSCs to an SC phenotype with typical repair characteristics and increase NGF secretion 11 . In addition, 3D SLC spheroids exhibited enhanced therapeutic potential both in vitro and after transplantation into a murine sciatic nerve injury model.…”

Section: Discussionmentioning

confidence: 99%

“…To induce differentiation of ADSCs into SLCs, ADSCs were cultured for three to five passages, and then 1 × 10 5 ADSCs/well were seeded onto collagen I-coated six-well plates (4810-010; IWAKI, Tokyo, Japan) for the conventional SLC group, or 2 × 10 4 ADSCs/well mixed with 0.2 mg/ml μ-piece (16629004; Fuji Film) were seeded onto ultra-low attachment 96-well plates (174925; Thermo Fisher Scientific) for the 3D SLC group. After cells in the 3D group had formed a spheroid in each well (approximately 24 h after seeding), our modified protocol with folic acid 11 was used to induce differentiation in both the conventional SLC and 3D SLC groups. All cells were incubated at 37 °C with 5% CO 2 in a humidified incubator.…”

Section: Methodsmentioning

confidence: 99%

“…However, because of limitations regarding the isolation, purification, and proliferation technology of primary SCs, there is an urgent need to develop a method to generate large numbers of replacement cells with low immunogenicity 10 . Our previous studies have successfully demonstrated a modified protocol to effectively induce Schwann-like cells (SLCs) from adipose-derived stem cells (ADSCs) into an SC phenotype with typical repair characteristics and increased secretion of nerve growth factor (NGF) 11 .…”

Section: Introductionmentioning

confidence: 99%

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Newly Generated 3D Schwann-Like Cell Spheroids From Human Adipose-Derived Stem Cells Using a Modified Protocol

et al. 2022

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Peripheral nerve injury (PNI) is a relatively frequent type of trauma that results in the suffering of many patients worldwide every year. Schwann cells (SCs) are expected to be applied in cell therapy because of their ability to promote peripheral nerve regeneration. However, the lack of clinically renewable sources of SCs hinders the application of SC-based therapies. Adipose-derived stem cells (ADSCs) have generated great interest in recent years because of their multipotency and ease of harvest, and they have already been verified to differentiate into Schwann-like cells (SLCs) in vitro. However, the efficiency of differentiation and the functions of SLCs remain unsatisfactory. We newly generated three-dimensional (3D) SLC spheroids from ADSCs using a modified protocol with human recombinant peptide (RCP) petaloid μ-piece. Morphological analysis, gene expression analysis by qRT-PCR, ELISA measurement of the secretion capabilities of neurotrophic factors, and neurite formation assay were performed to evaluate the functions of these 3D SLCs in vitro. Motor function recovery was measured in a sciatic nerve injury mouse model to analyze the nerve regeneration-promoting effect of 3D SLCs in vivo. The differentiation efficiency and the secretion of neurotrophic factors were enhanced in 3D SLCs compared with conventional SLCs. 3D SLCs could more effectively promote neurite growth and longer neurite extension in a neuron-like SH-SY5Y model. Additionally, 3D SLCs had a better therapeutic effect on nerve regeneration after transplantation into the sciatic nerve injury mouse model. These findings demonstrated that the potential of ADSC-derived SLCs to promote nerve regeneration could be significantly increased using our modified differentiation protocol and by assembling cells into a 3D sphere conformation. Therefore, these cells have great potential and can be used in the clinical treatment of PNI.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Newly Generated 3D Schwann-Like Cell Spheroids From Human Adipose-Derived Stem Cells Using a Modified Protocol

et al. 2022

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“…Exploring the mechanism of differentiation of BMSCs into SCLCs is the key to realizing stem cell‐based targeted therapy for PNI. Currently, most studies have focused on a single induction regimen and the morphological changes of SCLCs after induction, but the comparison of the induction regimen and studies of the mechanism of stem cell differentiation into SCLCs are rarely reported (Chen et al., 2021; Hörner et al., 2022). This study compared SCLCs induced by two distinct protocols based on various aspects, such as cell morphology, surface marker expression, apoptosis/necrosis, and proliferation.…”

Section: Discussionmentioning

confidence: 99%

Exploring miR‐21 as a key regulator in two distinct approaches of bone marrow stromal cells differentiation into Schwann‐like cells

Liu,

Wang,

Wei

et al. 2024

Synapse

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The differentiation of bone marrow stromal cells (BMSCs) into Schwann‐like cells (SCLCs) has the potential to promote the structural and functional restoration of injured axons. However, the optimal induction protocol and its underlying mechanisms remain unclear. This study aimed to compare the effectiveness of different induction protocols in promoting the differentiation of rat BMSCs into SCLCs and to explore their potential mechanisms. BMSCs were induced using two distinct methods: a composite factor induction approach (Protocol‐1) and a conditioned culture medium induction approach (Protocol‐2). The expression of Schwann cells (SCs) marker proteins and neurotrophic factors (NTFs) in the differentiated cells was assessed. Cell proliferation and apoptosis were also measured. During induction, changes in miR‐21 and Sprouty RTK signaling antagonist 2 (SPRY2) mRNA were analyzed. Following the transfection of BMSCs with miR‐21 agomir or miR‐21 antagomir, induction was carried out using both protocols, and the expression of SPRY2, ERK1/2, and SCs marker proteins was examined. The results revealed that NTFs expression was higher in Protocol‐1, whereas SCs marker proteins expression did not significantly differ between the two groups. Compared to Protocol‐1, Protocol‐2 exhibited enhanced cell proliferation and fewer apoptotic and necrotic cells. Both protocols showed a negative correlation between miR‐21 and SPRY2 expression throughout the induction stages. After induction, the miR‐21 agomir group exhibited reduced SPRY2 expression, increased ERK1/2 expression, and significantly elevated expression of SCs marker proteins. This study demonstrates that Protocol‐1 yields higher NTFs expression, whereas Protocol‐2 results in stronger SCLCs proliferation. Upregulating miR‐21 suppresses SPRY2 expression, activates the ERK1/2 signaling pathway, and promotes BMSC differentiation into SCLCs.

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The phenotypic changes of Schwann cells promote the functional repair of nerve injury

Liao,

Chen,

Luo

et al. 2024

Neuropeptides

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Effective <i>in vitro</i> differentiation of adipose-derived stem cells into Schwann-like cells with folic acid supplementation

Cited by 6 publications

References 42 publications

Newly Generated 3D Schwann-Like Cell Spheroids From Human Adipose-Derived Stem Cells Using a Modified Protocol

Newly Generated 3D Schwann-Like Cell Spheroids From Human Adipose-Derived Stem Cells Using a Modified Protocol

Exploring miR‐21 as a key regulator in two distinct approaches of bone marrow stromal cells differentiation into Schwann‐like cells

The phenotypic changes of Schwann cells promote the functional repair of nerve injury

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