Analysis of different conditioned media secreted by limbal progenitor cells in the modulation of corneal healing

The S17 murine stromal cell line was infected with retroviral vectors encoding the v-src and c-src oncogenes and cells expressing high levels of either pp60v-src or pp60c-src were isolated. Long-term bone marrow cultures (LTBMCs) established with these different stromal cell lines showed that progenitor cells proliferated to a greater extent in cultures with stromal cells that over-expressed either c-src or v-src. An increase in the number of granulocytes, monocytes, and colony- forming units granulocyte-macrophage (CFU-GM) in the nonadherent cell population of LTBMCs prepared with S17/v-src or S17/c-src stromal cells was observed. Conditioned media from the S17/v-src and S17/src stromal cell lines stimulated the formation of CFU-GM in the absence of additional hematopoietic cell growth factors. Conditioned media from S17/v-src and S17/c-src stimulated proliferation of the granulocyte- macrophage colony-stimulating factor (GM-CSF)-responsive cell line FDCP-1 and this stimulation was inhibited by neutralizing antisera to murine GM-CSF. An increase in the concentration of GM-CSF was confirmed by enzyme-linked immunosorbent assay. No secretion of interleukin-1 alpha (IL-1 alpha) or tumor necrosis factor-alpha was detected by any of the stromal cell lines. There was no increase in the secretion of either CSF-1 or IL-6 by either S17/v-src or S17/c-src. The addition of 1 micrograms/mL monoclonal anti-GM-CSF antibody to LTBMCs caused a decrease in the number of nonadherent cells in cultures established with each of the different stromal cell lines. Northern blot analysis showed no difference in the level of GM-CSF RNA among the different stromal cell lines. These studies suggest that the increased proliferation of hematopoietic progenitor cells in LTBMCs with S17/v-src or S17/c-src cells may result from a posttranscriptional event that elevates production of GM-CSF by the S17/c-src and S17/v-src stromal cells.

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Abstract TP470: Stimulation of Blood Vessel Formation by Human Neural Stem/Progenitor Cells is Mediated by Secreted Material

Stroke ◽

10.1161/str.51.suppl_1.tp470 ◽

2020 ◽

Vol 51 (Suppl_1) ◽

Author(s):

Brenda Gutierrez ◽

Lisa A Flanagan

Keyword(s):

Progenitor Cells ◽

Extracellular Vesicles ◽

Functional Recovery ◽

Conditioned Media ◽

Cell Contact ◽

Vessel Formation ◽

Angiogenic Proteins ◽

Neural Stem Progenitor Cells ◽

Stimulation Of

Human neural stem/progenitor cells (hNSPCs) have the potential to widen the current narrow treatment window for stroke as they improve functional recovery in rodent stroke models when transplanted weeks after stroke. One aspect of the hNSPC-induced functional recovery is increased angiogenesis and neovascularization in the peri-infarct region. Our lab created a human cell in vitro model of vessel formation by seeding hNSPCs and human endothelial progenitor cells (hEPCs) in a 3D scaffold composed of salmon fibrinogen, laminin, and hyaluronic acid that mimics brain tissue properties. Using our in vitro neurovascular model, we tested the hypothesis that hNSPC-secreted material plays a role in the stimulation of vessel formation. Our RNA-Seq data show that hNSPCs express high levels of secreted pro-angiogenic proteins, such as growth factors, matrix molecules, and cytokines, but hNSPCs might also impact vessel formation by secretion of extracellular vesicles or cell-contact mediated mechanisms. In order to determine the effect of hNSPC-secreted material on vessel formation, mCherry-labeled hEPCs were seeded in 3D scaffolds alone, with CellTracker Green-labeled hNSPCs, or with hNSPC-conditioned media containing hNSPC-secreted soluble factors and extracellular vesicles, such as exosomes. Vessel formation was quantified using AngioTool to determine total vessel length, number of branch points, and vessel percentage area. We found an increase in vessel formation in the presence of hNSPCs and hNSPC-conditioned media compared to hEPCs alone. In conclusion, material secreted by hNSPCs can recapitulate the increase in vessel formation induced by hNSPCs themselves. In future studies, we will determine whether hNSPC-derived exosomes are important for promoting vessel formation as they have therapeutic potential without the limitations of cell therapy.

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CELL-FREE CONDITIONED MEDIA FROM ENDOTHELIAL PROGENITOR CELLS ENHANCES ENDOTHELIAL ANGIOGENESIS, MIGRATION AND SURVIVAL: PARACRINE IMPLICATIONS FOR CELL THERAPY IN PULMONARY ARTERIAL HYPERTENSION

Canadian Journal of Cardiology ◽

10.1016/j.cjca.2014.07.234 ◽

2014 ◽

Vol 30 (10) ◽

pp. S156

Author(s):

C. Suen ◽

Y. Deng ◽

D. Stewart

Keyword(s):

Pulmonary Arterial Hypertension ◽

Cell Therapy ◽

Arterial Hypertension ◽

Progenitor Cells ◽

Endothelial Progenitor Cells ◽

Conditioned Media ◽

Endothelial Progenitor ◽

Pulmonary Arterial

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Anti-aging Properties of Conditioned Media of Epidermal Progenitor Cells Derived from Mesenchymal Stem Cells

Dermatology and Therapy ◽

10.1007/s13555-018-0229-2 ◽

2018 ◽

Vol 8 (2) ◽

pp. 229-244 ◽

Cited By ~ 4

Author(s):

Su Ji Sohn ◽

Ji Min Yu ◽

Eun Young Lee ◽

You Jin Nam ◽

Jinwan Kim ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Progenitor Cells ◽

Conditioned Media ◽

Aging Properties

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Conditioned media from AICAR-treated skeletal muscle cells increases neuronal differentiation of adult neural progenitor cells

Neuropharmacology ◽

10.1016/j.neuropharm.2018.10.041 ◽

2019 ◽

Vol 145 ◽

pp. 123-130 ◽

Cited By ~ 9

Author(s):

Hyo Youl Moon ◽

Sahar Javadi ◽

Matthew Stremlau ◽

Kyeong Jin Yoon ◽

Benjamin Becker ◽

...

Keyword(s):

Skeletal Muscle ◽

Progenitor Cells ◽

Neuronal Differentiation ◽

Neural Progenitor Cells ◽

Muscle Cells ◽

Neural Progenitor ◽

Conditioned Media ◽

Skeletal Muscle Cells

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Over-expression of c-src or v-src in bone marrow stromal cells stimulates hematopoiesis in long-term bone marrow culture

Blood ◽

10.1182/blood.v80.12.3079.3079 ◽

1992 ◽

Vol 80 (12) ◽

pp. 3079-3089 ◽

Cited By ~ 2

Author(s):

J Mladenovic ◽

SM Anderson

Keyword(s):

Bone Marrow ◽

Cell Line ◽

Progenitor Cells ◽

Cell Lines ◽

Stromal Cells ◽

Stromal Cell ◽

Conditioned Media ◽

Enzyme Linked Immunosorbent Assay ◽

Gm Csf

Abstract The S17 murine stromal cell line was infected with retroviral vectors encoding the v-src and c-src oncogenes and cells expressing high levels of either pp60v-src or pp60c-src were isolated. Long-term bone marrow cultures (LTBMCs) established with these different stromal cell lines showed that progenitor cells proliferated to a greater extent in cultures with stromal cells that over-expressed either c-src or v-src. An increase in the number of granulocytes, monocytes, and colony- forming units granulocyte-macrophage (CFU-GM) in the nonadherent cell population of LTBMCs prepared with S17/v-src or S17/c-src stromal cells was observed. Conditioned media from the S17/v-src and S17/src stromal cell lines stimulated the formation of CFU-GM in the absence of additional hematopoietic cell growth factors. Conditioned media from S17/v-src and S17/c-src stimulated proliferation of the granulocyte- macrophage colony-stimulating factor (GM-CSF)-responsive cell line FDCP-1 and this stimulation was inhibited by neutralizing antisera to murine GM-CSF. An increase in the concentration of GM-CSF was confirmed by enzyme-linked immunosorbent assay. No secretion of interleukin-1 alpha (IL-1 alpha) or tumor necrosis factor-alpha was detected by any of the stromal cell lines. There was no increase in the secretion of either CSF-1 or IL-6 by either S17/v-src or S17/c-src. The addition of 1 micrograms/mL monoclonal anti-GM-CSF antibody to LTBMCs caused a decrease in the number of nonadherent cells in cultures established with each of the different stromal cell lines. Northern blot analysis showed no difference in the level of GM-CSF RNA among the different stromal cell lines. These studies suggest that the increased proliferation of hematopoietic progenitor cells in LTBMCs with S17/v-src or S17/c-src cells may result from a posttranscriptional event that elevates production of GM-CSF by the S17/c-src and S17/v-src stromal cells.

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Conditioned media from endothelial progenitor cells cultured in simulated microgravity promote angiogenesis and bone fracture healing

Stem Cell Research & Therapy ◽

10.1186/s13287-020-02074-y ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Lingchi Kong ◽

Yan Wang ◽

Haixing Wang ◽

Qi Pan ◽

Rongtai Zuo ◽

...

Keyword(s):

Fracture Healing ◽

Progenitor Cells ◽

Endothelial Progenitor Cells ◽

Bone Fracture ◽

Mechanical Test ◽

Human Peripheral Blood ◽

Mapk Signaling ◽

Fracture Repair ◽

Conditioned Media ◽

Endothelial Progenitor

Abstract Background Paracrine signaling from endothelial progenitor cells (EPCs) is beneficial for angiogenesis and thus promotes tissue regeneration. Microgravity (MG) environment is found to facilitate the functional potentials of various stem or progenitor cells. The present study aimed to elucidate the effects of MG on pro-angiogenic properties and fracture repair capacities of conditioned media (CM) from EPCs. Methods Human peripheral blood-derived EPCs were cultured under MG or normal gravity (NG) followed by analysis for angiogenic gene expression. Furthermore, the serum-free CM under MG (MG-CM) or NG (NG-CM) were collected, and their pro-angiogenic properties were examined in human umbilical vein endothelial cells (HUVECs). In order to investigate the effects of MG-CM on fracture healing, they were injected into the fracture gaps of rat models, and radiography, histology, and mechanical test were performed to evaluate neovascularization and fracture healing outcomes. Results MG upregulated the expression of hypoxia-induced factor-1α (HIF-1α) and endothelial nitric oxide synthase (eNOS) and promoted NO release. Comparing to NG-CM, MG-CM significantly facilitated the proliferation, migration, and angiogenesis of HUVECs through NO-induced activation of FAK/Erk1/2-MAPK signaling pathway. In addition, MG-CM were verified to improve angiogenic activities in fracture area in a rat tibial fracture model, accelerate fracture healing, and well restore the biomechanical properties of fracture bone superior to NG-CM. Conclusion These findings provided insight into the use of MG bioreactor to enhance the angiogenic properties of EPCs’ paracrine signals via HIF-1α/eNOS/NO axis, and the administration of MG-CM favored bone fracture repair. Graphical abstract

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Impact of mesenchymal stem cells derived conditioned media on neural progenitor cells

General Physiology and Biophysics ◽

10.4149/gpb_2021026 ◽

2021 ◽

Vol 40 (06) ◽

pp. 551-559

Author(s):

Filip Humenik ◽

Sonja Jego ◽

Lubica Hornakova ◽

Marcela Maloveska ◽

Alexandra Valencakova-Agyagosova ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Progenitor Cells ◽

Neural Progenitor Cells ◽

Neural Progenitor ◽

Conditioned Media

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Therapeutic Effects of hiPSC-Derived Glial and Neuronal Progenitor Cells-Conditioned Medium in Experimental Ischemic Stroke in Rats

International Journal of Molecular Sciences ◽

10.3390/ijms22094694 ◽

2021 ◽

Vol 22 (9) ◽

pp. 4694

Author(s):

Diana Salikhova ◽

Tatiana Bukharova ◽

Elvira Cherkashova ◽

Daria Namestnikova ◽

Georgy Leonov ◽

...

Keyword(s):

Progenitor Cells ◽

Conditioned Medium ◽

Progenitor Cell ◽

Inflammatory Cytokine ◽

Conditioned Media ◽

Therapeutic Effects ◽

Neuronal Progenitor Cells ◽

Neuronal Progenitor ◽

Glial Progenitor Cells ◽

Glial Progenitor

Transplantation of various types of stem cells as a possible therapy for stroke has been tested for years, and the results are promising. Recent investigations have shown that the administration of the conditioned media obtained after stem cell cultivation can also be effective in the therapy of the central nervous system pathology (hypothesis of their paracrine action). The aim of this study was to evaluate the therapeutic effects of the conditioned medium of hiPSC-derived glial and neuronal progenitor cells in the rat middle cerebral artery occlusion model of the ischemic stroke. Secretory activity of the cultured neuronal and glial progenitor cells was evaluated by proteomic and immunosorbent-based approaches. Therapeutic effects were assessed by overall survival, neurologic deficit and infarct volume dynamics, as well as by the end-point values of the apoptosis- and inflammation-related gene expression levels, the extent of microglia/macrophage infiltration and the numbers of formed blood vessels in the affected area of the brain. As a result, 31% of the protein species discovered in glial progenitor cells-conditioned medium and 45% in neuronal progenitor cells-conditioned medium were cell type specific. The glial progenitor cell-conditioned media showed a higher content of neurotrophins (BDNF, GDNF, CNTF and NGF). We showed that intra-arterial administration of glial progenitor cells-conditioned medium promoted a faster decrease in neurological deficit compared to the control group, reduced microglia/macrophage infiltration, reduced expression of pro-apoptotic gene Bax and pro-inflammatory cytokine gene Tnf, increased expression of anti-inflammatory cytokine genes (Il4, Il10, Il13) and promoted the formation of blood vessels within the damaged area. None of these effects were exerted by the neuronal progenitor cell-conditioned media. The results indicate pronounced cytoprotective, anti-inflammatory and angiogenic properties of soluble factors secreted by glial progenitor cells.

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