SAT-LB103 Glucose-Dependent Insulinotropic Polypeptide Promotes Proliferation, Inhibits Apoptosis and Modifies Adipogenesis in Human Omental Adipose-Derived Stem Cells

Abstract Increased visceral fat correlates with a high risk of morbidity and mortality from diabetes and other metabolic diseases. To cope with changes of nutritional status, the adipose tissue undergoes dynamic remodeling, during which adipose derived stem cells (ADSCs) participate through cell proliferation and adipogenic differentiation into mature adipocytes. Besides, beige adipocytes formation from ADSCs, to dissipate energy as heat in mitochondrial via uncoupling protein1 (UCP1) has been proved to improve energy expenditure. Thus, modifying adipose remodeling and promoting beige adipogenesis of ADSCs in visceral fat bring much metabolic benefits. Newly listed LY3298176, an agonist targeted on glucose-dependent insulinotropic polypeptide (GIP) /glucagon-like peptide-1 (GLP-1) receptor, shows outstanding effect of reducing glucose and weight. Due to superior efficacy in dual-target agonist to GLP-1 monotherapy, and the unknown role of GIP in human visceral adipose, we aimed to clarify GIP’s role in undifferentiated ADSCs in vivo. We selected cell model derived from abdominal omental adipose tissue by obtaining ADSCs via primary culture from patients, because of wide-distributed GIP receptors in fat, and the dominant role of abdominal fat in metabolism. Then the cells were allowed to proliferate, or differentiate into adipocytes in the differentiation medium (DM), with or without co-treated with GIP or GIP3-42 (GIP receptor antagonist), followed by subsequently measurement. CCK-8, EdU incorporation, and cell cycle analysis were conducted to assess cellular proliferation. Annexin V FITC/PI stain, TUNEL and cleaved caspase3 detection were performed to evaluate apoptosis. The related signaling pathway was measured by Western blot and the validation was conducted by using pathway inhibitors followed with the above proliferation and apoptosis analysis. Besides, at the early stage of adipogenesis, mitotic clonal expansion (MCE) was reflected by cell cycle detection. Western blot analysis, quantitative real time-PCR (qRT-PCR), and Oil Red O staining were performed to evaluate adipogenesis. We found that GIP facilitated ADSCs viability and DNA synthesis, accelerated cell cycle progress and reduced palmitate-induced apoptosis by promoting phosphorylation of ERK1/2, AKT, PKA and AMPK. We further confirmed that ADSCs after confluence underwent MCE once induced by DM. GIP also modified adipogenesis by accelerating MCE, upregulating core transcription factor (PPARγ and C/EBPα), increasing beige-related markers (UCP1, PGC1α, PRDM16, et al) while suppressing white-related genes (ZFP423 and TLE3). In summary, we illustrated the efficacies of GIP on proliferation, apoptosis and adipogenesis (especially the beige adipocyte formation) of ADSCs, providing evidence of the additional metabolic benefits of GIP/GLP-1 dual-target agonist over GLP-1 agonist monotherapy in vivo.

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Recellularization of decellularized adipose tissue-derived stem cells: role of the cell-secreted extracellular matrix in cellular differentiation

Biomaterials Science ◽

10.1039/c7bm00695k ◽

2018 ◽

Vol 6 (1) ◽

pp. 168-178 ◽

Cited By ~ 16

Author(s):

V. Guneta ◽

Z. Zhou ◽

N. S. Tan ◽

S. Sugii ◽

M. T. C. Wong ◽

...

Keyword(s):

Stem Cells ◽

Extracellular Matrix ◽

Adipose Tissue ◽

Cellular Differentiation ◽

Adipose Derived Stem Cells ◽

Cellular Fate

The extracellular matrix (ECM) plays an important role in cellular fate decisions as demonstrated by adipose-derived stem cells (ASCs).

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Rapid Magneto-Sonoporation of Adipose-Derived Cells

Materials ◽

10.3390/ma14174877 ◽

2021 ◽

Vol 14 (17) ◽

pp. 4877

Author(s):

Miriam Filippi ◽

Boris Dasen ◽

Arnaud Scherberich

Keyword(s):

Stem Cells ◽

Tissue Repair ◽

Morphological Changes ◽

Adipose Derived Stem Cells ◽

Resonance Imaging ◽

Membrane Resealing ◽

First Time ◽

Stimulation Of

By permeabilizing the cell membrane with ultrasound and facilitating the uptake of iron oxide nanoparticles, the magneto-sonoporation (MSP) technique can be used to instantaneously label transplantable cells (like stem cells) to be visualized via magnetic resonance imaging in vivo. However, the effects of MSP on cells are still largely unexplored. Here, we applied MSP to the widely applicable adipose-derived stem cells (ASCs) for the first time and investigated its effects on the biology of those cells. Upon optimization, MSP allowed us to achieve a consistent nanoparticle uptake (in the range of 10 pg/cell) and a complete membrane resealing in few minutes. Surprisingly, this treatment altered the metabolic activity of cells and induced their differentiation towards an osteoblastic profile, as demonstrated by an increased expression of osteogenic genes and morphological changes. Histological evidence of osteogenic tissue development was collected also in 3D hydrogel constructs. These results point to a novel role of MSP in remote biophysical stimulation of cells with focus application in bone tissue repair.

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Modeling Adipogenesis: Current and Future Perspective

Cells ◽

10.3390/cells9102326 ◽

2020 ◽

Vol 9 (10) ◽

pp. 2326 ◽

Cited By ~ 2

Author(s):

Hisham F. Bahmad ◽

Reem Daouk ◽

Joseph Azar ◽

Jiranuwat Sapudom ◽

Jeremy C. M. Teo ◽

...

Keyword(s):

Stem Cells ◽

Adipose Tissue ◽

Crucial Role ◽

Critical Role ◽

Treatment Modalities ◽

Future Perspective ◽

Adipose Derived Stem Cells ◽

Physiological Importance ◽

And Personalized Medicine

Adipose tissue is contemplated as a dynamic organ that plays key roles in the human body. Adipogenesis is the process by which adipocytes develop from adipose-derived stem cells to form the adipose tissue. Adipose-derived stem cells’ differentiation serves well beyond the simple goal of producing new adipocytes. Indeed, with the current immense biotechnological advances, the most critical role of adipose-derived stem cells remains their tremendous potential in the field of regenerative medicine. This review focuses on examining the physiological importance of adipogenesis, the current approaches that are employed to model this tightly controlled phenomenon, and the crucial role of adipogenesis in elucidating the pathophysiology and potential treatment modalities of human diseases. The future of adipogenesis is centered around its crucial role in regenerative and personalized medicine.

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Abstract 402: Human CD271-positive Adipose Derived Stem Cells are the Angiogenic Subset Decreased by Donor Insulin Resistance

Circulation Research ◽

10.1161/res.127.suppl_1.402 ◽

2020 ◽

Vol 127 (Suppl_1) ◽

Author(s):

Oto Inoue ◽

Soichiro Usui ◽

Kosei Yamaguchi ◽

Yusuke Takeda ◽

Chiaki Goten ◽

...

Keyword(s):

Insulin Resistance ◽

Stem Cells ◽

Adipose Tissue ◽

Cell Therapy ◽

Capillary Density ◽

Adipose Derived Stem Cells ◽

Knock Out ◽

Healthy Donors ◽

Knock Out Mice

Introduction: Type 2 diabetes mellitus (T2DM) is an important risk factor for cardiovascular diseases (CVDs). Recently, cell therapy using adipose-derived stem cells (ADSCs) has emerged as an attractive therapy for severe CVDs because of their angiogenic potentials. However, whether and how T2DM would impair human ADSC angiogenic capacity is still uncertain. We previously reported that CD31 - CD34 + CD271 + ADSCs (CD271 + ADSCs) were specifically decreased in adipose tissue of T2DM patients. Therefore, we aimed to investigate the angiogenic capacity of CD271 + ADSCs. Furthermore, we evaluate which patients’ parameters regard as T2DM would decrease the amount of CD271 + ADSCs. Methods and Results: Human CD45 - CD34 + CD31 - ADSCs were obtained from subcutaneous adipose tissue of healthy donors, separated into CD271 + and CD271 - subsets by FACS, and cultured. Both subsets of ADSCs were assessed gene expression profile by microarray. Microarray analysis and validation PCR elucidated that PI3K/Akt/mTOR pathway was significantly up-regulated in CD271 + ADSCs compared to in CD271 - ADSCs. ( p < 0.05). Then, we compared in vivo angiogenic capacity in xenograft experiments of nude mice subjected to hindlimb ischemia. Angiogenesis was evaluated histologically using perfused lectin (capillary density) at day 14. Cell therapy using CD271 + ADSCs demonstrated about 3-fold more lectin + capillaries compared to CD271 - ADSCs or PBS injection ( p < 0.005, n = 5 / group). Next, we established cultured ADSCs obtained from CD271 knock-out mice (KO-ADSCs) and compared their angiogenic capacity with those from WT mice. Consistently, KO-ADSCs demonstrated impaired in vivo angiogenic capacity ( p < 0.005, n = 5 / group). Finally, we collected 23 samples of adipose tissue obtained from CVD patients and evaluated the frequency of CD271 + ADSCs in CD45 - CD34 + CD31 - ADSCs. Among studied parameters, HOMA-IR, an index of insulin resistance, was negatively correlated with the frequency of CD271+ ADSCs ( r = -0.64, p < 0.005). Conclusions: Human CD271 + ADSCs demonstrated enhanced in vivo angiogenic capacity with higher mTOR expression. Donor insulin resistance might decrease this regenerative subset of ADSCs. These findings would be critical for development and improvement of ADSC therapy.

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Adipocyte-Derived Adiponectin Positively Regulates Exit from Quiescence of Hematopoietic Stem Cells By Potentiating mTORC1 Activation after Myelotoxic Injury

Blood ◽

10.1182/blood.v126.23.777.777 ◽

2015 ◽

Vol 126 (23) ◽

pp. 777-777 ◽

Cited By ~ 1

Author(s):

Yosuke Masamoto ◽

Shunya Arai ◽

Tomohiko Sato ◽

Iseki Takamoto ◽

Naoto Kubota ◽

...

Keyword(s):

Cell Cycle ◽

Stem Cells ◽

Adipose Tissue ◽

Steady State ◽

Hematopoietic Stem Cells ◽

Hematopoietic Stem ◽

Hematopoietic Recovery ◽

Mtorc1 Activation

Abstract Myelotoxic injury unlocks the vigorous power of hematopoietic stem cells (HSCs) to replenish the hematopoietic system, making quiescent HSCs enter the cell cycle. Microscopically, it is well known that adipose tissue replaces cellular components in bone marrow (BM) after myeloablation by chemotherapeutic agents or irradiation. In a steady-state hematopoiesis, both HSC-intrinsic and -extrinsic mechanisms enforce quiescence of HSCs, and the interaction between HSCs and BM microenvironment has been drawing much attention in maintenance of the quiescence. In this meaning, it is supposed that the drastic change in BM microenvironment by myeloablation might trigger and promote the cell cycle entry of HSCs. We have previously reported that adiponectin, adipocyte-derived anti-diabetic hormone, indirectly enhances proliferation of murine immature myeloid progenitors upon granulocyte-colony stimulating factor treatment (emergency granulopoiesis) in Socs3-Stat3 dependent fashion by suppressing TNF-α production from macrophages (ASH meeting 2013, Abstract 221), however, its direct effect against HSCs in vivo is needed to be elucidated. Additionally, we have shown that both genetic loss and high-fat diet-induced reduction of adiponectin have no impact on steady-state hematopoiesis. Considering BM adipose tissue is an endocrine organ and adipocytes are the major cellular component in ablated marrow, we hypothesized that adiponectin derived from adipocytes might be implicated in HSC activation and subsequent hematopoietic recovery. Adiponectin-null (adipo-/-) mice showed significantly delayed hematopoietic recovery after 5-fluorouracil (5-FU) administration. In 5-FU-treated BM, adipo-/- SLAM-HSCs (CD150+ CD48- Lin- Sca-1+) and CD34- SLAM-HSCs were more quiescent than adipo+/+ counterparts. Adipo-/- mice survived longer than adipo+/+ control mice after serial 5-FU treatment. Taken into account our previous data showing that impaired emergency granulopoiesis of adipo-/- mice is Socs3-Stat3 dependent and Socs3 haploinsufficiency ameliorated the defect, we further investigated whether activation of adipo-/- HSCs on 5-FU treatment was potentiated by genetic loss of Socs3. But Socs3 haploinsufficiency had no capacity to revert impaired activation of adipo-/- HSCs, suggesting some mechanisms other than that of impaired emergency granulopoiesis in adipo-/- mice. Strikingly, adipo-/- HSCs were shown to be defective in mTORC1 activation, phosphorylation of S6 and mitochondrial activity after 5-FU treatment. In vivo rapamycin treatment cancelled the effect of adiponectin upon HSC activation by 5-FU, suggesting that adiponectin enhances HSC activation through mTORC1-dependent mechanism. Physiological isoform of adiponectin (full-length adiponectin) enhanced not only 5-FU-induced mTORC1 activation in vivo but also cytokine-induced activation in vitro, shortened the time to first division, without affecting subsequent proliferation of HSCs, in contrast to the previous report using non-physiological isoform (globular adiponectin) in vitro. The concentration of adiponectin in BM had a 4-fold increase after 5-FU treatment while the level in plasma remained unchanged. In a steady state, adiponectin level in adipocyte-rich tibia is higher than femur, suggesting local production of adiponectin constitutes a significant portion of BM adiponectin. Every BM cell components examined expressed adiponectin mRNA, and adipocytes had the highest. As 5-FU treatment had little effect on adiponectin expression in adipocytes, it was suggested that increased adipocytes in BM contributed to increased adiponectin upon myelotoxic injury. Furthermore, reciprocal transplants with adipo+/+ and adipo-/- mice demonstrated that adiponectin from BM environment of recipient mice plays a major role in the activation of HSC after in vivo 5-FU treatment and in vitro cytokine stimulation. These data reveal that adiponectin, produced mainly from increased adipocytes after myelotoxic injury, positively regulates HSC activation and subsequent hematopoietic recovery. Our data also highlight adipocytes as an essential source of adiponectin to ensure the proliferative burst of hematopoietic cells in myeloablated marrow. Adiponectin treatment could be clinically applied to relieve myelosuppression by chemotherapy. Disclosures No relevant conflicts of interest to declare.

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Immunomodulatory Effects of Adipose-Derived Stem Cells: Fact or Fiction?

BioMed Research International ◽

10.1155/2013/383685 ◽

2013 ◽

Vol 2013 ◽

pp. 1-8 ◽

Cited By ~ 53

Author(s):

Angelo A. Leto Barone ◽

Saami Khalifian ◽

W. P. Andrew Lee ◽

Gerald Brandacher

Keyword(s):

Stem Cells ◽

Immune Response ◽

Clinical Trials ◽

Immune System ◽

Solid Organ ◽

Adipose Derived Stem Cells ◽

Adipose Derived Stromal Cells

Adipose-derived stromal cells (ASCs) are often referred to as adipose-derived stem cells due to their potential to undergo multilineage differentiation. Their promising role in tissue engineering and ability to modulate the immune system are the focus of extensive research. A number of clinical trials using ASCs are currently underway to better understand the role of such cell niche in enhancing or suppressing the immune response. If governable, such immunoregulatory role would find application in several conditions in which an immune response is present (i.e., autoimmune conditions) or feared (i.e., solid organ or reconstructive transplantation). Although allogeneic ASCs have been shown to prevent acute GvHD in both preclinical and clinical studies, their potential warrants further investigation. Well-designed and standardized clinical trials are necessary to prove the role of ASCs in the treatment of immune disorders or prevention of tissue rejection. In this paper we analyze the current literature on the role of ASCs in immunomodulationin vitroandin vivoand discuss their potential in regulating the immune system in the context of transplantation.

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Macrophage-derived Apoptotic Vesicles Regulate Fate Commitment of Mesenchymal Stem Cells via miR155

10.21203/rs.3.rs-1101843/v1 ◽

2021 ◽

Author(s):

Yuan Zhu ◽

Xiao Zhang ◽

Kunkun Yang ◽

Yuzi Shao ◽

Ranli Gu ◽

...

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Western Blot ◽

Molecular Mechanisms ◽

Proliferation Ability ◽

Immunomodulatory Function

Abstract Background In tissue engineering, mesenchymal stem cells (MSCs) are common seed cells because of abundant sources, strong proliferation ability and immunomodulatory function. Numerous researches have demonstrated that MSC-macrophage crosstalk played a key role in the tissue engineering. Macrophages could regulate the differentiation of MSCs via different molecular mechanisms, including extracellular vesicles. Apoptotic macrophages could generate large amounts of apoptotic vesicles (apoVs), whereas the functions of macrophage-derived apoVs remain largely unknown. There was no research to clarify the role of macrophage-derived apoVs in MSC fate choices. In this study, we aimed to characterize macrophage-derived apoVs, and investigate the roles of macrophage-derived apoVs in the fate commitment of MSCs. Methods We characterized macrophage-derived apoVs, and investigated their role in MSC osteogenesis and adipogenesis in vitro and in vivo. Furthermore, we performed microRNA loss- and gain- of function experiments and western blot to determine the molecular mechanism. Results We found that macrophage-derived apoVs inhibited osteogenesis and promoted adipogenesis in vitro and in vivo. In mechanism, apoVs regulated osteogenesis and adipogenesis of MSCs by delivering microRNA155 (miR155). Conclusions Macrophage-derived apoVs could regulate the osteogenesis and adipogenesis of MSCs through delivering miR155, which provided novel insights for MSC-mediated tissue engineering.

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401 MIGRATION AND THERAPEUTIC POTENTIAL OF PORCINE ADULT ADIPOSE-DERIVED MESENCHYMAL STEM CELLS

Reproduction Fertility and Development ◽

10.1071/rdv22n1ab401 ◽

2010 ◽

Vol 22 (1) ◽

pp. 357 ◽

Cited By ~ 2

Author(s):

S. M. Wilson ◽

E. Monaco ◽

M. S. Goldwasser ◽

S. G. Clark ◽

W. L. Hurley ◽

...

Keyword(s):

Stem Cells ◽

Adipose Tissue ◽

Subcutaneous Adipose Tissue ◽

Adult Stem Cells ◽

Bone Defects ◽

Adipose Derived Stem Cells ◽

Subcutaneous Adipose ◽

Time Point

Bone marrow is one current source of adult stem cells for therapeutic purposes; however, the magnitude and accessibility of subcutaneous adipose tissue in humans make it an attractive alternative. Numerous in vitro studies have been conducted to determine how these cells act in vitro, but it is imperative to determine the vast abilities of these cells in vivo. The objective of this study was to evaluate in vivo migration and bone healing ability after transplanting adipose-derived stem cells (ADSC) in a swine model. Adipose-derived stem cells were isolated from subcutaneous adipose tissue of adult Yorkshire pigs and cultured in vitro. At 80 to 90% confluence/passage 3, the cells were trypsinized and labeled in suspension with carboxyfluorescein succinimidyl ester (CFDA-SE). This project included 20 pigs weighing between 63.5 and 81.7 kg. Bilateral mandibular osteoectomies with 10-mm defects were performed on each pig. Of the 20 pigs, half received a treatment of 2.5 million CFDA-SE labeled stem cells administered directly into each defect (DI), and the remaining half received a treatment of approximately 5 million CFDA-SE labeled stem cells through an ear vein injection via catheter (EVI). The time points were 1 h and 2 and 4 wk, with 2 pigs per time with the DI and EVI treatments. Pigs were slaughtered at each time, and spleen, liver, lung, kidney, ear vein, blood, and mandible tissues were collected. Blood samples were collected from the jugular vein with EDTA and processed via flow cytometry after collection. Tissues were fixed in 10% buffered formalin for histology. Fluorescent microscopy (CFDA-SE excitation/emission is 492/517 nm) has confirmed that transplanted ADSC do indeed migrate to a site of injury or trauma. Labeled cells were also present in blood collected from the 1-h time point group. Currently, we have not seen the presence of labeled ADSC in the other tissues (spleen, liver, lung, and kidney) after the 1-h time point. We did observe that ADSC administered by DI and EVI were able to significantly heal and regenerate bone defects within 4 wk post-surgery (P < 0.05, ANOVA with F-test), in contrast to bone defects in pigs that did not receive cell injections (control). Evidence of ADSC-related healing and bone regeneration was evident by gross visualization, dual-energy x-ray absorptiometry (DXA) and micro computer tomography (microCT) analysis. The clinical implications of these results are significant for treating many diseases in which inflammation or defects exist, such as cardiac disease, neurological disease, or traumatic injuries to both soft and hard tissue. If the adult stem cells can be harvested from fat, encouraged to produce bone or cartilage, and then reinserted into defects, treatment protocols for trauma victims could be developed that would reduce the need for alternate harvesting techniques for bone. This work was support by a grant from the Illinois Regenerative Medicine Institute (IDPH # 63080017).

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Role of Fzd6 in Regulating the Osteogenic Differentiation of Adipose-derived Stem Cells in Osteoporosis

10.21203/rs.3.rs-328417/v1 ◽

2021 ◽

Author(s):

Tianli Wu ◽

Zhihao Yao ◽

Gang Tao ◽

Fangzhi Lou ◽

Hui Tang ◽

...

Keyword(s):

Stem Cells ◽

Osteogenic Differentiation ◽

Wnt Signaling Pathway ◽

Osteogenic Potential ◽

Adipose Derived Stem Cells ◽

Alizarin Red ◽

Differentiation Potential

Abstract Objective: Although it has been demonstrated that adipose-derived stem cells (ASCs) from osteoporosis mice (OP-ASCs) exhibit impaired osteogenic differentiation potential, the molecular mechanism has not yet been elucidated. We found that Fzd6 was decreased in OP-ASCs compared with ASCs. This study investigates the effects and underlying mechanisms of Fzd6 in the osteogenic potential of OP-ASCs. Methods: Fzd6 expression in ASCs and OP-ASCs was measured by PCR gene chip. Fzd6 overexpression and silencing lentiviruses were used to evaluate the role of Fzd6 in the osteogenic differentiation of OP-ASCs. Real-time PCR (qPCR) and western blotting (WB) was performed to detect the expression of Fzd6 and bone-related molecules, including runt-related transcription factor 2 (Runx2) and osteopontin (Opn). Alizarin red staining and Alkaline phosphatase (ALP) staining was performed following osteogenic induction. Microscopic CT (Micro-CT), hematoxylin and eosin staining (H&E) staining, and Masson staining were used to assess the role of Fzd6 in osteogenic differentiation of osteoporosis (OP) mice in vivo.Results: Expression of Fzd6 was decreased significantly in OP-ASCs. Fzd6 silencing down-regulated the osteogenic ability of OP-ASCs in vitro. Overexpression of Fzd6 rescued the impaired osteogenic capacity in OP-ASCs in vitro. We obtained similar results in vivo.Conclusions: Fzd6 plays an important role in regulating the osteogenic ability of OP-ASCs both in vivo and in vitro. Overexpression of Fzd6 associated with the Wnt signaling pathway promotes the osteogenic ability of OP-ASCs, which provides new insights for the prevention and treatment of OP.

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