Limits...
Regulation of adipose tissue stromal cells behaviors by endogenic Oct4 expression control.

Kim JH, Jee MK, Lee SY, Han TH, Kim BS, Kang KS, Kang SK - PLoS ONE (2009)

Bottom Line: Exogenic Oct4 introduced a transdifferentiation priority into the neural lineage than mesodermal lineage.Global gene expression analysis results showed that Oct4 regulated target genes which could be characterized as differentially regulated genes such as pluripotency markers NANOG, SOX2, and KLF4 and markers of undifferentiated stem cells FOXD1, CDC2, and EPHB1.The negatively regulated genes included FAS, TNFR, COL6A1, JAM2, FOXQ1, FOXO1, NESTIN, SMAD3, SLIT3, DKK1, WNT5A, BMP1, and GLIS3 which are implicated in differentiation processes as well as a number of novel genes.

View Article: PubMed Central - PubMed

Affiliation: Department of Veterinary Biotechnology, Seoul National University, Seoul, Republic of Korea.

ABSTRACT

Background: To clarify the role of the POU domain transcription factor Oct4 in Adipose Tissue Stromal Cells (ATSCs), we investigated the regulation of Oct4 expression and other embryonic genes in fully differentiated cells, in addition to identifying expression at the gene and protein levels. The ATSCs and several immature cells were routinely expressing Oct4 protein before and after differentiating into specific lineages.

Methodology/principal findings and conclusions: Here, we demonstrated the role of Oct4 in ATSCs on cell proliferation and differentiation. Exogenous Oct4 improves adult ATSCs cell proliferation and differentiation potencies through epigenetic reprogramming of stemness genes such as Oct4, Nanog, Sox2, and Rex1. Oct4 directly or indirectly induces ATSCs reprogramming along with the activation of JAK/STAT3 and ERK1/2. Exogenic Oct4 introduced a transdifferentiation priority into the neural lineage than mesodermal lineage. Global gene expression analysis results showed that Oct4 regulated target genes which could be characterized as differentially regulated genes such as pluripotency markers NANOG, SOX2, and KLF4 and markers of undifferentiated stem cells FOXD1, CDC2, and EPHB1. The negatively regulated genes included FAS, TNFR, COL6A1, JAM2, FOXQ1, FOXO1, NESTIN, SMAD3, SLIT3, DKK1, WNT5A, BMP1, and GLIS3 which are implicated in differentiation processes as well as a number of novel genes. Finally we have demonstrated the therapeutic utility of Oct4/ATSCs were introduced into the mouse traumatic brain, engrafted cells was more effectively induces regeneration activity with high therapeutic modality than that of control ATSCs. Engrafted Oct4/ATSCs efficiently migrated and transdifferentiated into action potential carrying, functionally neurons in the hippocampus and promoting the amelioration of lesion cavities.

Show MeSH

Related in: MedlinePlus

The therapeutic efficacy of engrafted Oct4/ATSCs in a mouse model of a traumatic brain injury.(A) High neuroregenerative potency of Oct4/ATSCs in injured lesion of traumatic mouse brain. The arrows indicate engrafted and functionally transdifferentiated neurons in the hippocampus. (B) Electrophysiological properties of transdifferentiated neuron from engrafted Oct4/ATSCs from a lesion of traumatic injured brain slice. Whole-cell patch-clamp recordings of engrafted Oct4/ATSCs in traumatic brain slice. The presence of the ionic currents was first investigated by applying voltage ramps between 120 and 120 mV to the cells patch clamped using K-based pipette solution. ATSCs control engrafted brain slice does not display inward current, whereas Oct4/ATSCs engrafted brain slice actively produced a current. This current was approximately 75% blocked by 30 nM tetrodotoxin (TTX), although 300 nM TTX blocked it completely. Oct4/ATSCs engrafted brain slice express voltage-gated Na channels. Voltage-clamp recording at70 mV in extracellular solution containing 3 mM Mg2. Traces show spontaneous slow and fast currents that indicate that this transplanted Oct4/ATSCs receives synaptic contacts from host cells. This current was approximately 98% blocked by CNQX AP5 solution. (C) The Oct4/ATSCs engrafted traumatic brain demonstrated that there was a decrease in the lesion cavity size.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2747014&req=5

pone-0007166-g007: The therapeutic efficacy of engrafted Oct4/ATSCs in a mouse model of a traumatic brain injury.(A) High neuroregenerative potency of Oct4/ATSCs in injured lesion of traumatic mouse brain. The arrows indicate engrafted and functionally transdifferentiated neurons in the hippocampus. (B) Electrophysiological properties of transdifferentiated neuron from engrafted Oct4/ATSCs from a lesion of traumatic injured brain slice. Whole-cell patch-clamp recordings of engrafted Oct4/ATSCs in traumatic brain slice. The presence of the ionic currents was first investigated by applying voltage ramps between 120 and 120 mV to the cells patch clamped using K-based pipette solution. ATSCs control engrafted brain slice does not display inward current, whereas Oct4/ATSCs engrafted brain slice actively produced a current. This current was approximately 75% blocked by 30 nM tetrodotoxin (TTX), although 300 nM TTX blocked it completely. Oct4/ATSCs engrafted brain slice express voltage-gated Na channels. Voltage-clamp recording at70 mV in extracellular solution containing 3 mM Mg2. Traces show spontaneous slow and fast currents that indicate that this transplanted Oct4/ATSCs receives synaptic contacts from host cells. This current was approximately 98% blocked by CNQX AP5 solution. (C) The Oct4/ATSCs engrafted traumatic brain demonstrated that there was a decrease in the lesion cavity size.

Mentions: To determine the regenerative activity of Oct4/ATSCs for in vivo brain trauma, we evaluated the neurogenic potency through transdifferentiation of the Oct4/ATSCs. After neural induction, Oct4/ATSCs differentiated into neural cells expressed higher levels of TuJ than in differentiated control ATSCs (Fig. 7A). Extremely down-regulated Nestin expression was observed in Oct4-induced neural cells after induction (data not showed). Moreover, in in vivo lesion sites of injured brains, TuJ-positive neuron subpopulations generally survived or actively trans-differentiated into neurons after Oct4/ATSC engraftment (Fig. 7A). Immunohistochemical analyses of brains with traumatic injuries revealed that a large population of differentiated neural cells from Oct4/ATSCs displayed morphological and phenotypical characteristics of neurons (TuJ; approximately 35–40% of the total population) with highly improved rotor-rod activity (data not showed).


Regulation of adipose tissue stromal cells behaviors by endogenic Oct4 expression control.

Kim JH, Jee MK, Lee SY, Han TH, Kim BS, Kang KS, Kang SK - PLoS ONE (2009)

The therapeutic efficacy of engrafted Oct4/ATSCs in a mouse model of a traumatic brain injury.(A) High neuroregenerative potency of Oct4/ATSCs in injured lesion of traumatic mouse brain. The arrows indicate engrafted and functionally transdifferentiated neurons in the hippocampus. (B) Electrophysiological properties of transdifferentiated neuron from engrafted Oct4/ATSCs from a lesion of traumatic injured brain slice. Whole-cell patch-clamp recordings of engrafted Oct4/ATSCs in traumatic brain slice. The presence of the ionic currents was first investigated by applying voltage ramps between 120 and 120 mV to the cells patch clamped using K-based pipette solution. ATSCs control engrafted brain slice does not display inward current, whereas Oct4/ATSCs engrafted brain slice actively produced a current. This current was approximately 75% blocked by 30 nM tetrodotoxin (TTX), although 300 nM TTX blocked it completely. Oct4/ATSCs engrafted brain slice express voltage-gated Na channels. Voltage-clamp recording at70 mV in extracellular solution containing 3 mM Mg2. Traces show spontaneous slow and fast currents that indicate that this transplanted Oct4/ATSCs receives synaptic contacts from host cells. This current was approximately 98% blocked by CNQX AP5 solution. (C) The Oct4/ATSCs engrafted traumatic brain demonstrated that there was a decrease in the lesion cavity size.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2747014&req=5

pone-0007166-g007: The therapeutic efficacy of engrafted Oct4/ATSCs in a mouse model of a traumatic brain injury.(A) High neuroregenerative potency of Oct4/ATSCs in injured lesion of traumatic mouse brain. The arrows indicate engrafted and functionally transdifferentiated neurons in the hippocampus. (B) Electrophysiological properties of transdifferentiated neuron from engrafted Oct4/ATSCs from a lesion of traumatic injured brain slice. Whole-cell patch-clamp recordings of engrafted Oct4/ATSCs in traumatic brain slice. The presence of the ionic currents was first investigated by applying voltage ramps between 120 and 120 mV to the cells patch clamped using K-based pipette solution. ATSCs control engrafted brain slice does not display inward current, whereas Oct4/ATSCs engrafted brain slice actively produced a current. This current was approximately 75% blocked by 30 nM tetrodotoxin (TTX), although 300 nM TTX blocked it completely. Oct4/ATSCs engrafted brain slice express voltage-gated Na channels. Voltage-clamp recording at70 mV in extracellular solution containing 3 mM Mg2. Traces show spontaneous slow and fast currents that indicate that this transplanted Oct4/ATSCs receives synaptic contacts from host cells. This current was approximately 98% blocked by CNQX AP5 solution. (C) The Oct4/ATSCs engrafted traumatic brain demonstrated that there was a decrease in the lesion cavity size.
Mentions: To determine the regenerative activity of Oct4/ATSCs for in vivo brain trauma, we evaluated the neurogenic potency through transdifferentiation of the Oct4/ATSCs. After neural induction, Oct4/ATSCs differentiated into neural cells expressed higher levels of TuJ than in differentiated control ATSCs (Fig. 7A). Extremely down-regulated Nestin expression was observed in Oct4-induced neural cells after induction (data not showed). Moreover, in in vivo lesion sites of injured brains, TuJ-positive neuron subpopulations generally survived or actively trans-differentiated into neurons after Oct4/ATSC engraftment (Fig. 7A). Immunohistochemical analyses of brains with traumatic injuries revealed that a large population of differentiated neural cells from Oct4/ATSCs displayed morphological and phenotypical characteristics of neurons (TuJ; approximately 35–40% of the total population) with highly improved rotor-rod activity (data not showed).

Bottom Line: Exogenic Oct4 introduced a transdifferentiation priority into the neural lineage than mesodermal lineage.Global gene expression analysis results showed that Oct4 regulated target genes which could be characterized as differentially regulated genes such as pluripotency markers NANOG, SOX2, and KLF4 and markers of undifferentiated stem cells FOXD1, CDC2, and EPHB1.The negatively regulated genes included FAS, TNFR, COL6A1, JAM2, FOXQ1, FOXO1, NESTIN, SMAD3, SLIT3, DKK1, WNT5A, BMP1, and GLIS3 which are implicated in differentiation processes as well as a number of novel genes.

View Article: PubMed Central - PubMed

Affiliation: Department of Veterinary Biotechnology, Seoul National University, Seoul, Republic of Korea.

ABSTRACT

Background: To clarify the role of the POU domain transcription factor Oct4 in Adipose Tissue Stromal Cells (ATSCs), we investigated the regulation of Oct4 expression and other embryonic genes in fully differentiated cells, in addition to identifying expression at the gene and protein levels. The ATSCs and several immature cells were routinely expressing Oct4 protein before and after differentiating into specific lineages.

Methodology/principal findings and conclusions: Here, we demonstrated the role of Oct4 in ATSCs on cell proliferation and differentiation. Exogenous Oct4 improves adult ATSCs cell proliferation and differentiation potencies through epigenetic reprogramming of stemness genes such as Oct4, Nanog, Sox2, and Rex1. Oct4 directly or indirectly induces ATSCs reprogramming along with the activation of JAK/STAT3 and ERK1/2. Exogenic Oct4 introduced a transdifferentiation priority into the neural lineage than mesodermal lineage. Global gene expression analysis results showed that Oct4 regulated target genes which could be characterized as differentially regulated genes such as pluripotency markers NANOG, SOX2, and KLF4 and markers of undifferentiated stem cells FOXD1, CDC2, and EPHB1. The negatively regulated genes included FAS, TNFR, COL6A1, JAM2, FOXQ1, FOXO1, NESTIN, SMAD3, SLIT3, DKK1, WNT5A, BMP1, and GLIS3 which are implicated in differentiation processes as well as a number of novel genes. Finally we have demonstrated the therapeutic utility of Oct4/ATSCs were introduced into the mouse traumatic brain, engrafted cells was more effectively induces regeneration activity with high therapeutic modality than that of control ATSCs. Engrafted Oct4/ATSCs efficiently migrated and transdifferentiated into action potential carrying, functionally neurons in the hippocampus and promoting the amelioration of lesion cavities.

Show MeSH
Related in: MedlinePlus