Neuron-like differentiation of adipose tissue-derived stromal cells and vascular smooth muscle cells

doi:10.1111/j.1432-0436.2006.00081.x

Differentiation

Volume 74, Issues 9–10, December 2006, Pages 510-518

https://doi.org/10.1111/j.1432-0436.2006.00081.x Get rights and content

Abstract

Adipose tissue-derived stromal cells (ADSC) have previously been shown to possess stem cell properties such as transdifferentiation and self-renewal. Because future clinical applications are likely to use these adult stem cells in an autologous fashion, we wished to establish and characterize rat ADSC for pre-clinical tests. In the present study, we showed that rat ADSC expressed stem cell markers CD34 and STRO-1 at passage 1 but only STRO-1 at passage 3. These cells could also be induced to differentiate into adipocytes, smooth muscle cells, and neuron-like cells, the latter of which expressed neuronal markers S100, nestin, and NF70. Isobutylmethylxanthine (IBMX), indomethacin (INDO), and insulin were the active ingredients in a previously established neural induction medium (NIM); however, here we showed that IBMX alone was as effective as NIM in the induction of morphological changes as well as neuronal marker expression. Finally, we showed that vascular smooth muscle cells could also be induced by either NIM or IBMX to differentiate into neuron-like cells that expressed NF70.

References (57)

H. Betre et al.
Chondrocytic differentiation of human adipose-derived adult stem cells in elastin-like polypeptide
Biomaterials
(2006)
M. Brzoska et al.
Epithelial differentiation of human adipose tissue-derived adult stem cells
Biochem Biophys Res Commun
(2005)
Y. Cao et al.
Human adipose tissue-derived stem cells differentiate into endothelial cells in vitro and improve postnatal neovascularization in vivo
Biochem Biophys Res Commun
(2005)
W. Deng et al.
In vitro differentiation of human marrow stromal cells into early progenitors of neural cells by conditions that increase intracellular cyclic AMP
Biochem Biophys Res Commun
(2001)
A. Dicker et al.
Functional studies of mesenchymal stem cells derived from adult human adipose tissue
Exp Cell Res
(2005)
G.R. Erickson et al.
Chondrogenic potential of adipose tissue-derived stromal cells in vitro and in vivo
Biochem Biophys Res Commun
(2002)
J. Fujimura et al.
Neural differentiation of adipose-derived stem cells isolated from GFP transgenic mice
Biochem Biophys Res Commun
(2005)
K.G. Gaustad et al.
Differentiation of human adipose tissue stem cells using extracts of rat cardiomyocytes
Biochem Biophys Res Commun
(2004)
G.S. Jack et al.
Processed lipoaspirate cells for tissue engineering of the lower urinary tract: implications for the treatment of stress urinary incontinence and bladder reconstruction
J Urol
(2005)
S.K. Kang et al.
Improvement of neurological deficits by intracerebral transplantation of human adipose tissue-derived stromal cells after cerebral ischemia in rats
Exp Neurol
(2003)

J. Lu et al.

Adult bone marrow cells differentiate into neural phenotypes and improve functional recovery in rats following traumatic brain injury

Neurosci Lett

(2006)

G.F. Muschler et al.

Age- and gender-related changes in the cellularity of human bone marrow and the prevalence of osteoblastic progenitors

J Orthop Res

(2001)

A.M. Rodriguez et al.

The human adipose tissue is a source of multipotent stem cells

Biochimie

(2005)

A.M. Rodriguez et al.

Adipocyte differentiation of multipotent cells established from human adipose tissue

Biochem Biophys Res Commun

(2004)

K.M. Safford et al.

Neurogenic differentiation of murine and human adipose-derived stromal cells

Biochem Biophys Res Commun

(2002)

K.M. Safford et al.

Characterization of neuronal/glial differentiation of murine adipose-derived adult stromal cells

Exp Neurol

(2004)

J. Sanchez-Ramos et al.

Adult bone marrow stromal cells differentiate into neural cells in vitro

Exp Neurol

(2000)

M.J. Seo et al.

Differentiation of human adipose stromal cells into hepatic lineage in vitro and in vivo

Biochem Biophys Res Commun

(2005)

H.Y. Song et al.

Oncostatin M induces proliferation of human adipose tissue-derived mesenchymal stem cells

Int J Biochem Cell Biol

(2005)

B.M. Strem et al.

Expression of cardiomyocytic markers on adipose tissue-derived cells in a murine model of acute myocardial injury

Cytotherapy

(2005)

W. Wagner et al.

Comparative characteristics of mesenchymal stem cells from human bone marrow, adipose tissue, and umbilical cord blood

Exp Hematol

(2005)

M.M. Yaghoobi et al.

Nucleostemin, a coordinator of self-renewal, is expressed in rat marrow stromal cells and turns off after induction of neural differentiation

Neurosci Lett

(2005)

M. Yang et al.

In vitro and in vivo induction of bone formation based on ex vivo gene therapy using rat adipose-derived adult stem cells expressing BMP-7

Cytotherapy

(2005)

P.H. Ashjian et al.

In vitro differentiation of human processed lipoaspirate cells into early neural progenitors

Plast Reconstr Surg

(2003)

A. Banfi et al.

Bone marrow stromal damage after chemo/radiotherapy: occurrence, consequences and possibilities of treatment

Leuk Lymphoma

(2001)

A. Banfi et al.

High-dose chemotherapy shows a dose-dependent toxicity to bone marrow osteoprogenitors: a mechanism for post-bone marrow transplantation osteopenia

Cancer

(2001)

D. Bochinski et al.

The effect of neural embryonic stem cell therapy in a rat model of cavernosal nerve injury

BJU Int

(2004)

J. Case et al.

Clonal multilineage differentiation of murine common pluripotent stem cells isolated from skeletal muscle and adipose stromal cells

Ann NY Acad Sci

(2005)

Cited by (160)

Protein S-nitrosylation is involved in valproic acid-promoted neuronal differentiation of adipose tissue-derived stem cells
2024, Nitric Oxide - Biology and Chemistry
Neuronal differentiation of adipose tissue-derived stem cells (ASCs) is greatly promoted by valproic acid (VPA) with cAMP elevating agents thorough NO signaling pathways, but its mechanism is not fully understood. In the present study, we investigate the involvement of protein S-nitrosylation in the VPA-promoted neuronal differentiation of ASCs. The whole amount of S-nitrosylated protein was increased by the treatment with VPA alone for three days in ASCs. An inhibitor of thioredoxin reductase (TrxR), auranofin, further increased the amount of S-nitrosylated protein and enhances the VPA-promoted neuronal differentiation in ASCs. On the contrary, another inhibitor of TrxR, dinitrochlorobenzene, inhibited the VPA-promoted neuronal differentiation in ASCs even with cAMP elevating agents, which was accompanied by unexpectedly decreased S-nitrosylated protein. It was considered from these results that increased protein S-nitrosylation is involved in VPA-promoted neuronal differentiation of ASCs. By the proteomic analysis of S-nitrosylated protein in VPA-treated ASCs, no identified proteins could be specifically related to VPA-promoted neuronal differentiation. The identified proteins, however, included those involved in the metabolism of substances regulating neuronal differentiation, such as aspartate and glutamate.
Hydrogen sulfide potently promotes neuronal differentiation of adipose tissue-derived stem cells involving nitric oxide-mediated signaling cascade with the aid of cAMP-elevating agents
2022, Nitric Oxide - Biology and Chemistry
Neuronal differentiation of adipose tissue-derived stem cells (ASCs) is potently promoted by valproic acid (VPA) through a gaseous signaling molecule, nitric oxide (NO). Here, we investigated the involvement of hydrogen sulfide (H₂S), another gaseous signaling molecule, in neuronal differentiation of ASCs. VPA-promoted neuronal differentiation of ASCs was accompanied by increased intracellular H₂S and sulfane sulfur with increased mRNA expression of enzymes synthesizing sulfane sulfur including cystathionine β-synthase (CBS), of which inhibition reduced the differentiation efficiency. H₂S donors, GYY4137 (GYY) or NaHS, potently promoted neuronal differentiation of ASCs when cAMP-elevating agents, dibutyryl cyclic adenosine monophosphate and isobutyl methyl-xanthine, were added as neuronal induction medium (NIM). Neuronal differentiation of ASCs promoted by NaHS or GYY was accompanied by Ca²⁺ entry and increased mRNA expression of voltage-gated Ca²⁺ channels. NaHS or GYY also increased mRNA expression of enzymes of the NO-citrulline cycle including inducible NO synthase (iNOS). It was concluded from these results that H₂S potently promoted differentiation of ASCs into neuronal cells expressing functional voltage-gated Ca²⁺ channels with the aid of cAMP-elevating agents, involving NO-mediated signaling cascade. These effects of H₂S were also considered as a partial mechanism for the VPA-promoted neuronal differentiation of ASCs.
Valproic acid promotes differentiation of adipose tissue-derived stem cells to neuronal cells selectively expressing functional N-type voltage-gated Ca<sup>2+</sup> channels
2022, Biochemical and Biophysical Research Communications
The differentiation of adipose tissue-derived stem cells (ASCs) to neuronal cells is greatly promoted by valproic acid (VPA), and is synergistically enhanced by the following treatment with neuronal induction medium (NIM) containing cAMP-elevating agents. In the present study, we investigated the synergism between VPA and NIM in neuronal differentiation of ASCs, assessed by the expression of neurofilament medium polypeptide (NeFM), with respect to Ca²⁺ entry. VPA (2 mM) treatment for 3 days followed by NIM for 2 h synergistically increased the incidence of neuronal cells differentiated from ASCs to an extent more than VPA alone treatment for 6 days, shortening the time required for the differentiation. VPA increased intracellular Ca²⁺ and the mRNAs of voltage-gated Ca²⁺ channels, Cacna1b (Cav2.2) and Cacna1h (Cav3.2), in ASCs. Inward currents through Ca²⁺ channels were evoked electrophysiologically at high voltage potential in ASCs treated with VPA. NIM reduced the mRNAs of NeFM and Cacna1b in VPA-promoted neuronal differentiation of ASCs. It was concluded that functional N-type voltage-gated Ca²⁺ channels (Cav2.2) are selectively expressed in VPA-promoted neuronal differentiation of ASCs. NIM seems to enhance the mRNA translation of molecules required for the differentiation. Neuronal cells obtained from ASCs by this protocol will be used as a cell source for regenerative therapy of neurological disorders associated with altered Cav2.2 activity.
Valproic acid up-regulates the whole NO-citrulline cycle for potent iNOS-NO signaling to promote neuronal differentiation of adipose tissue-derived stem cells
2021, Nitric Oxide - Biology and Chemistry
Valproic acid (VPA) remarkably promotes the differentiation of adipose tissue-derived stem cells (ASCs) to mature neuronal cells through nitric oxide (NO) signaling due to up-regulated inducible NO synthase (iNOS) as early as within 3 days. Here, we investigated mechanisms of VPA-promoted neuronal differentiation of ASCs concerning the NO-citrulline cycle, the metabolic cycle producing NO. Cultured rat ASCs were differentiated to mature neuronal cells rich in dendrites and expressing a neuronal marker by treatments with VPA at 2 mM for 3 days and subsequently with the neuronal induction medium for 2 h. Inhibitor (α-methyl-d, l-aspartic acid, MDLA) of arginosuccinate synthase (ASS), a key enzyme of the NO-citrulline cycle, abolishes intracellular NO increase and VPA-promoted neuronal differentiation in ASCs. l-Arginine, the substrate of iNOS, restores the promotion effect of VPA, being against MDLA. Immunocytochemistry showed that ASS and iNOS were increased in ASCs expressing neurofilament medium polypeptide (NeFM), a neuronal marker, by VPA and NIM synergistically. Real-time RT-PCR analysis showed that mRNAs of Ass and arginosuccinate lyase (Asl) in the NO-citrulline cycle were increased by VPA. Chromatin immunoprecipitation assay indicated that Ass and Asl were up-regulated by VPA through the acetylation of their associated histone. From these results, it was considered that VPA up-regulated the whole NO-citrulline cycle, which enabled continuous NO production by iNOS in large amounts for potent iNOS-NO signaling to promote neuronal differentiation of ASCs. This may also indicate a mechanism enabling short-lived NO to function conveniently as a potent signaling molecule that can disappear quickly after its role.
Valproic acid promotes mature neuronal differentiation of adipose tissue-derived stem cells through iNOS–NO–sGC signaling pathway
2019, Nitric Oxide - Biology and Chemistry
Valproic acid (VPA) remarkably promotes the differentiation of adipose tissue-derived stem cells (ASCs) to mature neuronal cells, enabling neuronal induction within only three days. Here, we investigated the involvement of NO-signaling in the VPA-promoted neuronal differentiation of ASCs as a possible mechanism. Cultured rat ASCs were differentiated to matured neuronal cells rich in dendrites and expressing βIII-tubulin protein, a neuronal marker, by treatments with VPA at 2 mM for 3 days and subsequently with the neuronal induction medium (NIM) containing cAMP-elevating agents for 2 h. Increased intracellular NO was detected in neuronal cells differentiated from ASCs treated with VPA by a fluorescence NO-specific probe, diaminofluorescein-FM diacetate. However, a NO donor (NOC18) increased the incidence of neuronal cells only to a lesser extent than VPA, indicating the insufficiency of exogenous NO. RT-PCR analysis of ASCs treated with VPA showed increased mRNA expression of inducible nitric oxide synthase (iNOS) with the acetylation of its associated histone H3K9. iNOS inhibitors (1400 W and dexamethasone) or a soluble guanylate cyclase (sGC) inhibitor (ODQ) decreased the incidence of neuronal cells differentiated from ASCs treated with VPA. These inhibitors also decreased the mRNA expression of mature neuronal markers, neurofilament medium polypeptide (NeFM) and microtubule-associated protein 2 (MAP2), as well as βIII-tubulin (TUBB3), to various extents. It was considered from these results that VPA promoted mature neuronal differentiation of ASCs through the iNOS–NO–sGC signaling pathway. This provided insights into the regulated neuronal differentiation of ASCs in clinical applications.
CD34+ synovial fibroblasts exhibit high osteogenic potential in synovial chondromatosis
2024, Cell and Tissue Research

View all citing articles on Scopus

View full text

ORIGINAL ARTICLENeuron-like differentiation of adipose tissue-derived stromal cells and vascular smooth muscle cells

Abstract

Biomaterials

Biochem Biophys Res Commun

Biochem Biophys Res Commun

Biochem Biophys Res Commun

Exp Cell Res

Biochem Biophys Res Commun

Biochem Biophys Res Commun

Biochem Biophys Res Commun

J Urol

Exp Neurol

Neurosci Lett

J Orthop Res

Biochimie

Biochem Biophys Res Commun

Biochem Biophys Res Commun

Exp Neurol

Exp Neurol

Biochem Biophys Res Commun

Int J Biochem Cell Biol

Cytotherapy

Exp Hematol

Neurosci Lett

Cytotherapy

In vitro differentiation of human processed lipoaspirate cells into early neural progenitors

Plast Reconstr Surg

Bone marrow stromal damage after chemo/radiotherapy: occurrence, consequences and possibilities of treatment

Leuk Lymphoma

High-dose chemotherapy shows a dose-dependent toxicity to bone marrow osteoprogenitors: a mechanism for post-bone marrow transplantation osteopenia

Cancer

The effect of neural embryonic stem cell therapy in a rat model of cavernosal nerve injury

BJU Int

Clonal multilineage differentiation of murine common pluripotent stem cells isolated from skeletal muscle and adipose stromal cells

Ann NY Acad Sci

ORIGINAL ARTICLE
Neuron-like differentiation of adipose tissue-derived stromal cells and vascular smooth muscle cells