Icariside II Promotes the Differentiation of Adipose Tissue-Derived Stem Cells to Schwann Cells to Preserve Erectile Function after Cavernous Nerve Injury

Zheng, Tao,Zhang, Tian-biao,Wang, Chao-liang,Zhang, Wei-xing,Jia, Dong-hui,Yang, Fan,Sun, Yang-yang,Ding, Xiao-ju,Wang, Rui Korean Society for Molecular and Cellular Biology 2018 Molecules and cells Vol.41 No.6

Icariside II (ICA II) is used in erectile dysfunction treatment. Adipose tissue-derived stem cells (ADSCs) are efficient at improving erectile function. This study aimed to explore the action mechanism of ADSCs in improving erectile function. ADSCs were isolated from the adipose tissues of rats. Cell proliferation was determined using the Cell Counting Kit-8 (CCK-8) assay. The expressions of mRNA and protein were determined separately through qRT-PCR and western blot. The endogenous expressions of related genes were regulated using recombinant plasmids and cell transfection. A Dual-Luciferase Reporter Assay was performed to determine the interaction between miR-34a and STAT3. Rat models with bilateral cavernous nerve injuries (BCNIs) were used to assess erectile function through the detection of mean arterial pressure (MAP) and intracavernosal pressure (ICP). ICA II promoted ADSCs' proliferation and differentiation to Schwann cells (SCs) through the inhibition of miR-34a. Suppressed miR-34a promoted the differentiation of ADSCs to SCs by upregulating STAT3. ICA II promoted the differentiation of ADSCs to SCs through the miR-34a/STAT3 pathway. The combination of ICA II and ADSCs preserved the erectile function of the BCNI model rats. ADSCs treated with ICA II markedly preserved the erectile function of the BCNI model rats, which was reversed through miR-34a overexpression. ICA II promotes the differentiation of ADSCs to SCs through the miR34a/STAT3 pathway, contributing to erectile function preservation after the occurrence of a cavernous nerve injury.

Icariside II Promotes the Differentiation of Adipose Tissue-Derived Stem Cells to Schwann Cells to Preserve Erectile Function after Cavernous Nerve Injury

Tao Zheng,Tian-biao Zhang,Chao-liang Wang,Wei-xing Zhang,Dong-hui Jia,Fan Yang,Yang-yang Sun,Xiao-ju Ding,Rui Wang 한국분자세포생물학회 2018 Molecules and cells Vol.41 No.6

Icariside II (ICA II) is used in erectile dysfunction treatment. Adipose tissue-derived stem cells (ADSCs) are efficient at improving erectile function. This study aimed to explore the action mechanism of ADSCs in improving erectile function. ADSCs were isolated from the adipose tissues of rats. Cell proliferation was determined using the Cell Counting Kit-8 (CCK-8) assay. The expressions of mRNA and protein were determined separately through qRT-PCR and western blot. The endogenous expressions of related genes were regulated using recombinant plasmids and cell transfection. A Dual-Luciferase Reporter Assay was performed to determine the interaction between miR-34a and STAT3. Rat models with bilateral cavernous nerve injuries (BCNIs) were used to assess erectile function through the detection of mean arterial pressure (MAP) and intracavernosal pressure (ICP). ICA II promoted ADSCs’ proliferation and differentiation to Schwann cells (SCs) through the inhibition of miR-34a. Suppressed miR-34a promoted the differentiation of ADSCs to SCs by up-regulating STAT3. ICA II promoted the differentiation of ADSCs to SCs through the miR-34a/STAT3 pathway. The combination of ICA II and ADSCs preserved the erectile function of the BCNI model rats. ADSCs treated with ICA II markedly preserved the erectile function of the BCNI model rats, which was reversed through miR-34a overexpression. ICA II pro-motes the differentiation of ADSCs to SCs through the miR-34a/STAT3 pathway, contributing to erectile function preservation after the occurrence of a cavernous nerve injury.

Comparison of the Infant and Adult Adipose-Derived Mesenchymal Stem Cells in Proliferation, Senescence, Anti-oxidative Ability and Differentiation Potential

Wu Szu-Hsien,Yu Jin-Huei,Liao Yu-Ting,Liu Kuo-Hao,Chiang En-Rung,Chang Ming-Chau,Wang Jung-pan 한국조직공학과 재생의학회 2022 조직공학과 재생의학 Vol.19 No.3

BACKGROUND: Infant adipose-derived mesenchymal stem cells (ADSCs) collected from excised polydactyly fat tissue, which was surgical waste, could be cultured and expanded in vitro in this study. In addition, the collecting process would not cause pain in the host. In this study, the proliferation, reduction of senescence, anti-oxidative ability, and differentiation potential in the infant ADSCs were compared with those in the adult ADSCs harvested from thigh liposuction to determine the availability of infant ADSCs. METHODS: Proliferation was determined by detecting the fold changes in cell numbers and doubling time periods. Senescence was analyzed by investigating the age-related gene expression levels and the replicative stress. The superoxide dismutase (SOD) gene expression, adipogenic, neurogenic, osteogenic, and tenogenic differentiation were compared by RTqPCR. The chondrogenic differentiation efficiency was also determined using RT-qPCR and immunohistochemical staining. RESULTS: The proliferation, SOD (SOD1, SOD2 and SOD3) gene expression, the stemness-related gene (c-MYC) and telomerase reverse transcriptase of the infant ADSCs at early passages were enhanced compared with those of the adults’. Cellular senescence related genes, including p16, p21 and p53, and replicative stress were reduced in the infant ADSCs. The adipogenic genes (PPARc and LPL) and neurogenic genes (MAP2 and NEFH) of the infant ADSC differentiated cells were significantly higher than those of the adults’ while the expression of the osteogenic genes (OCN and RUNX) and tenogenic genes (TNC and COL3A1) of both demonstrated opposite results. The chondrogenic markers (SOX9, COL2 and COL10) were enhanced in the infant ADSC differentiated chondrogenic pellets, and the expression levels of SODs were decreased during the differentiation process. CONCLUSION: Cultured infant ADSCs demonstrate less cellular senescence and replicative stress, higher proliferation rates, better antioxidant defense activity, and higher potential of chondrogenic, adipogenic and neurogenic differentiation.

Advanced Properties of Urine Derived Stem Cells Compared to Adipose Tissue Derived Stem Cells in Terms of Cell Proliferation, Immune Modulation and Multi Differentiation

강혜숙,최석환,김범수,최재영,박강백,권태균,전소영 대한의학회 2015 Journal of Korean medical science Vol.30 No.12

Adipose tissue stem cells (ADSCs) would be an attractive autologous cell source. However, ADSCs require invasive procedures, and has potential complications. Recently, urine stem cells (USCs) have been proposed as an alternative stem cell source. In this study, we compared USCs and ADSCs collected from the same patients on stem cell characteristics and capacity to differentiate into various cell lineages to provide a useful guideline for selecting the appropriate type of cell source for use in clinical application. The urine samples were collected via urethral catheterization, and adipose tissue was obtained from subcutaneous fat tissue during elective laparoscopic kidney surgery from the same patient (n = 10). Both cells were plated for primary culture. Cell proliferation, colony formation, cell surface markers, immune modulation, chromosome stability and multi-lineage differentiation were analyzed for each USCs and ADSCs at cell passage 3, 5, and 7. USCs showed high cell proliferation rate, enhanced colony forming ability, strong positive for stem cell markers expression, high efficiency for inhibition of immune cell activation compared to ADSCs at cell passage 3, 5, and 7. In chromosome stability analysis, both cells showed normal karyotype through all passages. In analysis of multi-lineage capability, USCs showed higher myogenic, neurogenic, and endogenic differentiation rate, and lower osteogenic, adipogenic, and chondrogenic differentiation rate compared to ADSCs. Therefore, we expect that USC can be an alternative autologous stem cell source for muscle, neuron and endothelial tissue reconstruction instead of ADSCs.

Advanced Properties of Urine Derived Stem Cells Compared to Adipose Tissue Derived Stem Cells in Terms of Cell Proliferation, Immune Modulation and Multi Differentiation

Kang, Hye Suk,Choi, Seock Hwan,Kim, Bum Soo,Choi, Jae Young,Park, Gang-Baek,Kwon, Tae Gyun,Chun, So Young The Korean Academy of Medical Sciences 2015 JOURNAL OF KOREAN MEDICAL SCIENCE Vol.30 No.12

<P>Adipose tissue stem cells (ADSCs) would be an attractive autologous cell source. However, ADSCs require invasive procedures, and has potential complications. Recently, urine stem cells (USCs) have been proposed as an alternative stem cell source. In this study, we compared USCs and ADSCs collected from the same patients on stem cell characteristics and capacity to differentiate into various cell lineages to provide a useful guideline for selecting the appropriate type of cell source for use in clinical application. The urine samples were collected via urethral catheterization, and adipose tissue was obtained from subcutaneous fat tissue during elective laparoscopic kidney surgery from the same patient (n = 10). Both cells were plated for primary culture. Cell proliferation, colony formation, cell surface markers, immune modulation, chromosome stability and multi-lineage differentiation were analyzed for each USCs and ADSCs at cell passage 3, 5, and 7. USCs showed high cell proliferation rate, enhanced colony forming ability, strong positive for stem cell markers expression, high efficiency for inhibition of immune cell activation compared to ADSCs at cell passage 3, 5, and 7. In chromosome stability analysis, both cells showed normal karyotype through all passages. In analysis of multi-lineage capability, USCs showed higher myogenic, neurogenic, and endogenic differentiation rate, and lower osteogenic, adipogenic, and chondrogenic differentiation rate compared to ADSCs. Therefore, we expect that USC can be an alternative autologous stem cell source for muscle, neuron and endothelial tissue reconstruction instead of ADSCs.</P>

The Effect of Silicon Ion on Proliferation and Osteogenic Differentiation of Human ADSCs

( Ki Joo Kim ),( Young Joon Jeon ),( Jung Ho Lee ),( Sang Tae Ahn ),( Soo Hong Lee ),( Dong Woo Cho ),( Jong Won Rhie ) 한국조직공학과 재생의학회 2010 조직공학과 재생의학 Vol.7 No.2

Tissue engineering requires biopolymer scaffolds with compressive strength. Bioglass produces the compressive strength required of new scaffolds, and ionized Bioglass improves cell proliferation and differentiation. Ionized Bioglass contains phosphorous, sodium, calcium, and silicon ions, but the contributions of single ions or ion complexes to the cellular effects of Bioglass have not been determined. Here, we examined the effect of silicon on cell growth and differentiation. Culture medium containing 2, 4, 6, 8, and 10 μM silicon ions was produced in a chemical neutralization reaction by mixing a sodium silicate solution and hydroxychloride, and human adiposederived stromal cells (ADSCs) were cultured in the silicon-containing medium. Cell growth was followed for 9 daysin culture, using a Cell Counting Kit-8 assay. Osteogenic differentiation was examined by alizarin red S staining and RT-PCR for osteogenic genes at 10, 20, and 30 days of culture. The proliferation of human ADSCs cultured in medium containing 2 or 4 μM silicon was significantly greater than that of ADSCs in the control medium. Differentiation of human ADSCs was increased by supplementation of the differentiation medium with 2 μM silicon. This work demonstrates that 2 μM silicon accelerated proliferation and osteogenic differentiation of human ADSCs.

BDNF Enhancement of Neuronal Differentiation with Adipose-Derived Stem Cells(ADSCs)

( Chae Yoon Lim ),( Kwon Il Keun ),( Jeong June Choi ),( Hee Jin Lee ),( Sang Hee Park ),( Weon Sun Lee ),( Dong Hoon Kwak ),( Ji Youl Lee ),( Young Joon Jun ),( Won Bae Lee ),( Sung Eun Kim ),( Dai J 한국조직공학·재생의학회 2008 조직공학과 재생의학 Vol.5 No.4

Adipose-derived stem cells(ADSCs) are multipotent cells that can be manipulated to differentiate into different cell lines and are considered a potential cell source for regenerative medicine and tissue engineering. The brain derived neurotrophic factor (BDNF) is known to stimulate stem cells to differentiate into neuronal cells as well as promote plasticity. The purpose of this study was to evaluate the effects of BDNF on the neuronal differentiation of ADSCs. The proliferation of the ADSCs was increased by BDNF in a dose-dependent manner. The expression of neuronal markers including Nestin and microtubule-associated protein(MAP2) were highly upregulated by BDNF. In addition, BDNF considerably suppressed the expression of glial fibrillary acidic protein(GFAP), a preventative glial cell or astrocyte marker. Using immunocytochemistry, we confirmed the results by real time-PCR. An increased production of Nestin and MAP2 by BDNF, but not GFAP was observed. These data show that supplementation of the ADSCs with BDNF effectively maintained cell survival, proliferation and differentiation of these cells into neuronal precursor cells. In conclusion, specific neuronal differentiation of ADSCs after treatment with BDNF increased the generation of a mature neuronal phenotype.

지방유래 줄기세포가 이식된 Vicryl Mesh의 섬유화와 육아조직 형성에 미치는 영향

허찬영 ( Chan Yeong Heo ),하현정 ( Hyun Jung Ha ),김지회 ( Ji Hoe Kim ),정의철 ( Eui Cheol Jeong ),은석찬 ( Seok Chan Eun ),백롱민 ( Rong Min Baek ),김석화 ( Suk Wha Kim ),민경원 ( Kyung Won Minn ) 한국조직공학·재생의학회 2010 조직공학과 재생의학 Vol.7 No.1

Vicryl mesh consists of polyglactin 910, a synthetic absorbable copolymer made from 90% glycolide and 10% L-lactide. The empirical formula is (C2H2O2)m(C3H4O2)n. The mesh is knitted from uncoated and undyed fiber identical in composition to that used in Vicryl sutures, which has been found to be inert, nonantigenic, nonpyrogenic and to elicit only a mild tissue reaction during absorption. It may be used wherever temporary wound or organ support is required, particularly in instances in which compliant and stretchable support is desired. Successful use is reported for instance as support for the correction of breast deformity. Because a putative potential stem cell population within the adipose tissue has been found to possess therapeutic potentials, we authors sought to determine the effect of adipose tissue-derived stem cells(ADSCs) on Vicryl mesh implantation. Sprague-Dawley rats were divided into two groups one is control, given the surgery of simple Vicryl mesh implantation and the other is stem cell group, given the surgery of mesh implantation and ADSCs(N=12 for each group). Before the surgery ADSCs of stem cell group were isolated from the inguinal fat pads and expanded ex vivo for three passages. Control group received multi-folded Vicryl mesh implantation to the back of the rats and Stem cell group received the surgery of Vicryl mesh implantation and ADSCs. After four weeks of operation, matrix formation, capillary density and collagen production were assessed quantatively under the light microscope. ADSCs led to a statistically significant increase in fibrosis and mature scar formation compared with the control group in histologic examination (p < 0.05). ADSCs was found to be efficacious in promoting fibrosis and scar maturation leading volume replacement on experimental Vicryl mesh implantation. This study might be used as useful volume replacement with Vicryl mesh for correcting breast deformity after conservative surgery.

지방유래줄기세포의 지방분화과정에서 활성산소가 미치는 영향

장학,민경희,박영인,김요한,민경원 대한성형외과학회 2011 Archives of Plastic Surgery Vol.38 No.2

Purpose: Stem cells continue to receive research attention in the clinical fields, and adipose - derived stem cells (ADSCs) have been shown to be a good source raw material. Many plastic surgeons are researching the ADSC adipogenesis with a view to conducting clinical trials, and many attempts have been made to identify the factors that promote the adipogenesis of ADSCs, but comparatively few correlation studies have been undertaken to explore the relation between reactive oxygen species (ROS) and the ADSC adipogenesis. We undertook this study is to investigate the effects of ROS on ADSC adipogenesis. Methods: ADSCs were isolated and cultured from abdominal adipose tissue, and cultured in different media; 1) DMEM (control), 2) adipogenesis induction culture medium, 3) adipogenesis induction culture medium with ROS (20 μM / 50 μM H2O2), 4) adipogenesis induction culture medium containing ROS (20 μM / 50 μM H2O2) and antioxidant (10 μM / 20 μM Deferoxamine). We compared adipogenesis in these different media by taking absorbance measurements after Oil - Red O staining every 5 days. Results: After culturing for 20 days, significant differences were observed between these various culture groups. Absorbance results showed significantly more adipogenesis had occurred in media containing adipogenesis induction culture medium and H2O2(in a H2O2 dose - dependently manner) than in media containing adipogenesis induction culture medium and no H2O2 (p < 0.001). Furthermore, in media containing adipogenesis induction culture medium, H2O2, and antioxidant, absorbance results were significantly lower than in adipogenesis induction culture medium and H2O2 (p < 0.001). Conclusions: These findings suggest that ROS promote the adipogenesis of ADSCs. We suggests that ROS could be used in the adipose tissue engineering to improve fat cell differentiation and implantable fat tissue organization.

Osteogenic Differentiation of Stem Cells in a Gelatin-Hydroxyapatite Nanocomposite

( Dae Seok Hwang ),( Uk Kyu Kim ),( Myung Cheol Chang ),( Yong Deok Kim ),( Sang Hun Shin ),( In Kyo Chung ) 한국조직공학·재생의학회 2009 조직공학과 재생의학 Vol.6 No.12

Gelatin-hydroxyapatite (HG) nanocomposites have an inorganic nanostructure similar to that of bone. However, HG nanocomposites exhibit only osteoconductivity and not osteoinductivity. Recently, stem cell seeding on nanocomposite scaffolds has been attempted, in an effort to improve osteoinductivity. The present study examined the effects of gelatin and glutaraldehyde (GA) addition on the structural features of an HG nanocomposite scaffold containing stem cells. HG nanocomposites were fabricated by the precipitation of hydroxyapatite within a gelatin matrix. Four materials were synthesized as follows: HG3 (10 g HA in 3 g gelatin solution), HG3+GA (10 g HA in 3 g gelatin solution; GA addition), HG4 (10 g HA in 4 g gelatin solution), HG4+GA (10 g HA in 4 g gelatin solution; also termed GA addition). After fabrication, scaffolds were seeded with adipose tissue-derived stromal cells (ADSCs) obtained from rabbit suprascapular fat pads. After cell culture, osteogenic inductivity was tested by appropriate staining, scanning electron microscopy (SEM), colorimetric assays, biochemical tests for alkaline phosphatase (ALP), and an osteocalcin (OC) detection kit. Cell attachment to the scaffold surface was observed by SEM. Cell viability in scaffolds was quantified using the MTS assay. The maximum ALP and OC contents were seen in HG3-derived material. Substrates with added GA showed lower ALP and OC levels than did those without GA. The HG3 group without GA showed the best response with respect to attachment, proliferation, and osteogenic differentiation of ADSCs. These findings suggest that a HG nanocomposite scaffold with ADSCs may be osteoinductive.