Journal: Bioactive Materials

Article Title: Acellular nerve xenografts based on supercritical extraction technology for repairing long-distance sciatic nerve defects in rats

doi: 10.1016/j.bioactmat.2022.03.014

Figure Lengend Snippet: Evaluation of the cytotoxicity of the ANXs scaffold in vitro. (A, D) Living/dead double staining of Schwann cells grown on the ANXs scaffold for 3 days and 7 days (live: green, dead: red). (B, E) SEM images of Schwann cells growing on CD SD and CD + scCO 2 NG scaffolds for 7 days (the picture on the right is an enlarged view of the yellow area in the picture on the left). (C, F) Immunofluorescence images of Schwann cells growing on CD SD and CD + scCO 2 NG scaffolds for 7 days, respectively (S100: red, nucleus: blue). (G) Quantification of the number of live/dead double-stained Schwann cells in each region (0.36 mm 2 ). Data are presented as the mean ± SD (n = 3). (H) The CCK-8 assay was performed after 1, 3, 5 and 7 days of cell culture. Data are presented as the mean ± SD (n = 5). (I, J) Quantitative analysis of the GDNF and NGF expression levels of Schwann cells on the ANXs scaffold. Data are presented as the mean ± SD (n = 5). Statistical analysis: n.s. no significances, **p < 0.01, *p < 0.05.

Article Snippet: In brief, the medium of each group was centrifuged at 1500 rpm and 4 °C for 10 min, the concentration of NGF and BDNF in the supernatant was assessed using ELISA kits, the rat GDNF ELISA kit (EK0363, BOSTER, China) and the rat NGF/NGFβ ELISA kit (EK0471, BOSTER, China), and the absorbance of each well at 450 nm was determined using a spectrophotometer (EPOCH TAKE 3, Bio-Tek, USA).

Techniques: In Vitro, Double Staining, Immunofluorescence, Staining, CCK-8 Assay, Cell Culture, Expressing

Journal: Oncotarget

Article Title: Melatonin promotes goat spermatogonia stem cells (SSCs) proliferation by stimulating glial cell line-derived neurotrophic factor (GDNF) production in Sertoli cells

doi: 10.18632/oncotarget.12720

Figure Lengend Snippet: A. Light field of SSCs cultured with melatonin and GDNF, bar=50 μm. B. Cell density after being cultured with different cell mediums at 48 h. The initial number was 5*10 4 . C. QRT-PCR and western blot analysis of proliferation, self-renewal and Sertoli cell markers. D. Western blot analysis of proliferation and Sertoli cell markers. *, P<0.05,**, P<0.01.

Article Snippet: GDNF levels were determined by using a Human glial cell line-derived neurotrophic factor (GDNF) ELISA Kit (BOSTER).

Techniques: Cell Culture, Quantitative RT-PCR, Western Blot

Journal: Oncotarget

Article Title: Melatonin promotes goat spermatogonia stem cells (SSCs) proliferation by stimulating glial cell line-derived neurotrophic factor (GDNF) production in Sertoli cells

doi: 10.18632/oncotarget.12720

Figure Lengend Snippet: A. ELISA analysis of GDNF levels in the SSCs medium. B. Western Blot analysis of phosphorylation levels of AKT and ERK. *, P<0.05,**, P<0.01.

Article Snippet: GDNF levels were determined by using a Human glial cell line-derived neurotrophic factor (GDNF) ELISA Kit (BOSTER).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Phospho-proteomics

Journal: International Journal of Molecular Sciences

Article Title: De novo Neurosteroidogenesis in Human Microglia: Involvement of the 18 kDa Translocator Protein

doi: 10.3390/ijms22063115

Figure Lengend Snippet: Human microglia C20 and HC3 cells: time-dependence of the pregnenolone production and genotyping for TSPO rs6971 polymorphism. ( A , B ) C20 and HMC3 cell samples were incubated in serum-free saline medium at time zero in the presence of the inhibitors trilostane and SU10603, and a kinetic analysis of pregnenolone released from microglia cells was performed. After various incubation times, the saline medium was collected from distinct cell samples and pregnenolone content was quantified by indirect ELISA. As shown in the figure, pregnenolone released from C20 and HMC3 cells increased in a time-dependent manner. A single incubation time (120 min) was conducted for the “inhibitor-free” samples named 120 without Inhibitors (120 W/O In). In this case, pregnenolone was not present in the collected samples, suggesting the conversion of pregnenolone into other neurosteroids. Pregnenolone levels were normalized based on the number of cells evaluated after crystal violet staining. Data are presented as means ± SEMs of three independent experiments. Statistical analysis was determined by one-way ANOVA followed by Bonferroni’s post-test: * p < 0.05, ** p < 0.01, *** p < 0.001 vs. 60 min; **** p < 0.0001, 120 W/O In vs. 120 min. ( C ) Comparison of pregnenolone production between human microglial and U87MG cells. ( D ) The classical stimulus of peripheral steroidogenesis (cAMP pathway activation) did not promote neurosteroidogenesis in microglial cells. Indeed, pregnenolone production by microglial cells was not stimulated following treatment with the known adenylate cyclase activator forskolin. However, the starvation phase before forskolin treatment led to a high increase in pregnenolone production. Data are represented as means ± SEMs of two independent experiments. The significance of the differences was determined by one-way ANOVA, which was followed by Bonferroni’s post-test: ** p < 0.01, **** p < 0.0001 vs. control. ( E ) The C20 and HMC3 cell genotyping for TSPO rs6971 was performed by restriction fragment length polymorphism (RFLP) analysis. The amplification product (329 bp) derived from genomic DNA was subjected to digestion by the restriction endonuclease NruI. Following the enzymatic digestion, the samples were subjected to agarose gel electrophoresis. Only the amplification product containing an Ala147 allele can be digested by NruI and generates restriction fragments (184 and 145 bp). As shown in the figure, C20 and HMC3 cells exhibited the restriction pattern typical of the Ala147 homozygous genotype.

Article Snippet: Genomic DNA (gDNA) was extracted by using the “Genomic DNA extraction kit” (Thermo Fisher Scientific, Waltham, Massachusetts USA) from a cell pellet composed of 6 × 10 5 cells approximately, following the manufacturer’s protocol. gDNA was quantified with NanoDrop Instrument (Thermo Fisher Scientific) and used as a template for PCR reaction performed with Phusion Master Mix (Thermo Fisher Scientific).

Techniques: Incubation, Saline, Indirect ELISA, Staining, Comparison, Activation Assay, Control, Amplification, Derivative Assay, Agarose Gel Electrophoresis

Journal: Frontiers in Cellular Neuroscience

Article Title: Hypoxic culture of umbilical cord mesenchymal stem cell-derived sEVs prompts peripheral nerve injury repair

doi: 10.3389/fncel.2022.897224

Figure Lengend Snippet: Cellular localization and uptake of fluorescently labeled sEVs with SCs. (A) Cellular colocalization: SP8 was used to photograph sEVs prelabeled with PKH67 (green fluorescence) with Hoechst 33,342 (blue fluorescence)-stained SC nuclei. (B) Western blot: Detection of ERK1/2, ZEB2, and c-JUN expression levels in NC Schwann cells and after 48-h treatment with hypoxia sEVs. (C) Real-time PCR: Inflammatory and other restoration-related factors (IL-1β, IL-6, TNF-α, etc.) were detected. (D) ELISA test: GDNF NDF and NT-3 in the supernatant of Schwann cell preparation. Statistical significance, * p < 0.05,** p < 0.01, and **** p < 0.0001. *Significant difference.

Article Snippet: The following ELISA kits were: mouse glial cell line-derived neurotrophic factor (GDNF), ELISA kit (CSB-E07341m, CUSABIO), mouse neurotrophin 3 (NT-3), ELISA kit (CSB-E04687m, CUSABIO), and mouse nerve growth factor (NGF), ELISA kit (CSB-E04684m, CUSABIO).

Techniques: Labeling, Fluorescence, Staining, Western Blot, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Experimental Neurobiology

Article Title: Glial Cell Line-derived Neurotrophic Factor-overexpressing Human Neural Stem/Progenitor Cells Enhance Therapeutic Efficiency in Rat with Traumatic Spinal Cord Injury

doi: 10.5607/en.2019.28.6.679

Figure Lengend Snippet: Lists of antibodies used for immunostaining and primers used for qPCR

Article Snippet: The levels of human GDNF were measured using a human GDNF DuoSet ELISA Development kit (R&D systems) according to the manufacturer’s instructions.

Techniques: Immunostaining, Immunocytochemistry, Immunohistochemistry, Amplification, Sequencing

Journal: Experimental Neurobiology

Article Title: Glial Cell Line-derived Neurotrophic Factor-overexpressing Human Neural Stem/Progenitor Cells Enhance Therapeutic Efficiency in Rat with Traumatic Spinal Cord Injury

doi: 10.5607/en.2019.28.6.679

Figure Lengend Snippet: In vitro and in vivo characterization of GDNF-hNSPCs. (A) Compared with Mock-hNSPCs (DAPI + , blue), many of GDNF-hNSPCs express GDNF (green). (B) An ELISA of GDNF in Mock- and GDNF-CM. (C, D) Representative images of Mock- and GDNF-hNSPCs stained for differentiation markers and quantification of the percentages of cells that expressed these markers. DAPI + Cells (blue) express nestin (red), GFAP (green), Tuj1 (red), PDGFR (green), and O4 (red). (E, F) Representative images of CM-treated SH-SY5Y cells and quantification of average neurite length under different experimental conditions. (G) hNuc + GDNF–hNSPCs (red) implanted into the injured spinal cord show robust engraftment and an extensive distribution throughout the lesion and adjacent areas. The boxed area is shown at high magnification in the right panel. The asterisk indicates the lesion epicenter and arrows indicate the cell transplantation sites rostral and caudal to the lesion epicenter. (H, I) Representative images of Mock- and GDNF-hNSPCs stained for differentiation markers in vivo and quantification of the percentages of cells that expressed these markers. Confocal images show that hNSPCs positive for the human cell markers Ku80, STEM101, and STEM121 colocalize with differentiation markers (green). (J) Engrafted STEM121 + GDNF-hNSPCs (red) express GDNF (green). Scale bars: 50 μm in A, D; 100 μm in E; 1 mm in G; and 30 μm in I, J. Data represent the means±SEM. *p<0.05 vs. Mock-hNSPCs in C, H; *p<0.05 vs. vehicle, ***p<0.001 vs. vehicle, ##p<0.01 vs. Mock-hNSPCs in F.

Article Snippet: The levels of human GDNF were measured using a human GDNF DuoSet ELISA Development kit (R&D systems) according to the manufacturer’s instructions.

Techniques: In Vitro, In Vivo, Enzyme-linked Immunosorbent Assay, Staining, Transplantation Assay

Journal: Experimental Neurobiology

Article Title: Glial Cell Line-derived Neurotrophic Factor-overexpressing Human Neural Stem/Progenitor Cells Enhance Therapeutic Efficiency in Rat with Traumatic Spinal Cord Injury

doi: 10.5607/en.2019.28.6.679

Figure Lengend Snippet: Grafted GDNF-hNSPCs enhance neurite outgrowth, axonal extension, and myelination in the injured spinal cord. (A) Multiple NF + neuronal processes (green) extend over engrafted hNuc + human cells (red). Compared with Mock-hNSPCs, GDNF-hNSPCs significantly enhance neurite extension and host axonal sprouting. Arrows indicate hNuc + donor-derived cells (red) and arrowheads indicate NF + neuronal fibers (green). (B) Under high magnification, many NF + neuronal processes (red) are co-labeled with an anti-MBP antibody (green). Arrowheads indicate co-labeled myelinated neuronal fibers (yellow or orange). (C) The intensity of MBP staining in engrafted and adjacent areas is markedly higher in the GDNF-hNSPC-treated group than in the Mock-hNSPC-treated group. Arrows indicate hNuc + donor-derived cells (red) and arrowheads indicate MBP + myelinated neuronal fibers (green). (D) Representative images of LFB staining (blue) in the lesion and adjacent areas of the spinal cord. Dashed lines indicate demyelinated areas. (E) Quantification of the demyelinated volume in the different experimental groups. (F, G) Representative electron microscopic images of axially sectioned spinal cords (T 8 ~T 10 ) and quantification of G-ratio. (H, I) Representative images of Schwann/2E immunostaining (green) in the injured spinal cords and quantification of immunostaining density in the different experimental groups. Scale bars: 1 mm in A, C, D; 100 μm in B; 1 μm in F, and 500 μm in H. Data represent the means±SEM. *p<0.05 vs. vehicle, #p<0.01 vs. Mock-hNSPCs.

Article Snippet: The levels of human GDNF were measured using a human GDNF DuoSet ELISA Development kit (R&D systems) according to the manufacturer’s instructions.

Techniques: Derivative Assay, Labeling, Staining, Immunostaining

Journal: Experimental Neurobiology

Article Title: Glial Cell Line-derived Neurotrophic Factor-overexpressing Human Neural Stem/Progenitor Cells Enhance Therapeutic Efficiency in Rat with Traumatic Spinal Cord Injury

doi: 10.5607/en.2019.28.6.679

Figure Lengend Snippet: Grafted GDNF-hNSPCs protect the dCST, promote the formation of detour circuits, and reduce lesion volume and glia scar formation. (A) Representative images of BDA-labeled dCST fibers (red) at 6 mm rostral to the lesion epicenter in the different experimental groups. The boxed areas in the left panels are shown at high magnification in the right panels. (B) Quantification of the number of dCST fibers up to 6 mm rostral and caudal to the lesion epicenter. (C) Representative images of dCST collaterals (black, left panel) and FG-labeled PSNs (white, right panel) in the cervical enlargement of the spinal cord in the different experimental groups. The white dashed lines in the right panel indicate the margins between gray and white matter of the spinal cord. (D) Quantification of dCST collaterals and FG-labeled PSNs in the cervical enlargement. (E) Representative images of the contacts between dCST collaterals (black) and PSNs (brown) in the cervical enlargement. Arrowheads show closely located collateral fibers and PSNs. (F) Quantification of the number of contacts between dCST collaterals and PSNs. (G) Representative images of GFAP immunostaining in the lesion and adjacent areas of the spinal cord in the different experimental groups. Dashed lines indicate GFAP-negative areas. (H) Quantification of lesion volume, spared tissue volume, and glial scars. Scale bars: 100 μm in A, C; 40 μm in E; and 1 mm in G. Data represent the means±SEM. *p<0.05 vs. vehicle, ***p<0.001 vs. vehicle, #p<0.01 vs. Mock-hNSPCs.

Article Snippet: The levels of human GDNF were measured using a human GDNF DuoSet ELISA Development kit (R&D systems) according to the manufacturer’s instructions.

Techniques: Labeling, Immunostaining

Journal: Experimental Neurobiology

Article Title: Glial Cell Line-derived Neurotrophic Factor-overexpressing Human Neural Stem/Progenitor Cells Enhance Therapeutic Efficiency in Rat with Traumatic Spinal Cord Injury

doi: 10.5607/en.2019.28.6.679

Figure Lengend Snippet: Transplantation of GDNF-hNSPCs promotes motosensory recovery, reduces expression of voltage-gated sodium channels and NPY, and increases expression of GABA. (A) The BBB open-field walking scores before transplantation and at various time points post-transplantation. (B) Von Frey tests for mechanical allodynia in lesioned hindlimbs of rats with SCI before transplantation and at various time points post-transplantation. (C) qRT-PCR analysis of Nav1.3 and Nav1.9 expression in the T 8 ~T 10 segment of the spinal cord. (D, E) Representative images and quantification of NPY expression in lamina I of the dorsal horn in the T 8 segment of the spinal cord. (F, G) Representative images and quantification of GABA expression in the T 8 ~T 10 segment of the spinal cord. (H) Many engrafted hNuc + GDNF-hNSPCs (red, arrowheads) express GABA (green, arrowheads) adjacent endogenous GABA + cells (green, arrows). Scale bars: 200 μm in D; 500 μm in F; and 50 μm in H. Data represent the means±SEM. *p<0.05 vs. vehicle, **p<0.01 vs. vehicle, ***p<0.001 vs. vehicle, #p<0.05 vs. Mock-hNSPCs, ##p<0.01 vs. Mock-hNSPCs in A, B. *p<0.05 vs. vehicle, #p<0.05 vs. Mock-hNSPCs in C, E, and G.

Article Snippet: The levels of human GDNF were measured using a human GDNF DuoSet ELISA Development kit (R&D systems) according to the manufacturer’s instructions.

Techniques: Transplantation Assay, Expressing, Quantitative RT-PCR

Journal: The Journal of Physiology

Article Title: Cellular properties and chemosensory responses of the human carotid body

doi: 10.1113/jphysiol.2013.263657

Figure Lengend Snippet: A, GDNF mRNA expression in human CB samples from 66- and 17-year-old subjects. Note the lack of expression of GDNF in the SCG tissue (66-year-old subject). GAPDH was used as housekeeping gene to normalise mRNA. B, GDNF content (in picograms per microgram of total protein) measured by ELISA in human and rat CB and SCG. C, GDNF protein levels in CBs from subjects below (n= 8) and above (n= 5) 50 years of age. *P < 0.05 and **P < 0.01 (Mann–Whitney U test).

Article Snippet: This can be explained by the cross-reactivity of the ELISA GDNF detection kit with other structurally related trophic factors, especially with members of the TGF-β superfamily (for detailed information see the technical specifications of the GDNF Emax Immunoassay system, TB221, Promega).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Journal of Zhejiang University. Science. B

Article Title: Zirconium oxide ceramic foam: a promising supporting biomaterial for massive production of glial cell line-derived neurotrophic factor *

doi: 10.1631/jzus.B1400163

Figure Lengend Snippet: GDNF productivity in ZrO2 ceramic foam astrocyte culturing system

Article Snippet: ELISA assessment of GDNF production The supernatants from the astrocytes culturing system were collected for ELISA assay using a rat GDNF ELISA kit (Promega) according to the protocol provided by the manufacturer.

Techniques:

Journal: Current Research in Pharmacology and Drug Discovery

Article Title: Long-term, stable, targeted biodelivery and efficacy of GDNF from encapsulated cells in the rat and Goettingen miniature pig brain

doi: 10.1016/j.crphar.2020.04.001

Figure Lengend Snippet: (a) ELISA analysis of encapsulated clonal ARPE-19 ​cell lines demonstrates robust continued secretion of GDNF from clone-125. While GDNF secretion from clones 20 and 120 decreased between 4 and 8 weeks, secretion from clone-125 remained stable and even rose during the same period in vivo. (b) Immunohistochemistry of tissue sections at 8 weeks post implant confirms robust diffusion of GDNF from encapsulated clone-125 ​cells throughput the implanted striatum and overlying cortex. Scale bar, 500 ​μm in panel b and 75 ​μm in panel c. H&E staining on longitudinal sections of devices confirms excellent viability of encapsulated clone −125 ​cells. Clone-125 was used for all subsequent studies. Data are expressed as mean ​± ​SEM of 4 animals each with bilateral implants per group/time-point. ∗∗∗p<0.0001; Student's t test for unpaired data.

Article Snippet: A commercially available GDNF ELISA kit (DuoSet® for human GDNF R&D Systems, Minneapolis, MN) was used to quantify GDNF device secretion prior to implantation and again following retrieval from the brain.

Techniques: Enzyme-linked Immunosorbent Assay, Clone Assay, In Vivo, Immunohistochemistry, Diffusion-based Assay, Staining

Journal: Current Research in Pharmacology and Drug Discovery

Article Title: Long-term, stable, targeted biodelivery and efficacy of GDNF from encapsulated cells in the rat and Goettingen miniature pig brain

doi: 10.1016/j.crphar.2020.04.001

Figure Lengend Snippet: Long-term, sustained delivery of GDNF and resulting elevation tissue levels of GDNF within the implanted striatum. Fig. 2 a demonstrates that GDNF device secretion peaks early post implantation but then stabilizes and remains consistent for at least 6 months in vivo. Fig. 2 b confirms that GDNF delivery results in sustained tissue levels within the implanted striatum. When explanted from the striatum the output of the devices over time generally paralleled the enhanced tissue concentrations with an early peak in delivery followed by a longer, sustained output similar to that noted from the same devices immediately prior to implantation. All data are expressed as mean ​± ​SEM of 4 animals each with bilateral implants per time-point. BLQ ​= ​below limit of quantitation.

Article Snippet: A commercially available GDNF ELISA kit (DuoSet® for human GDNF R&D Systems, Minneapolis, MN) was used to quantify GDNF device secretion prior to implantation and again following retrieval from the brain.

Techniques: In Vivo, Quantitation Assay

Journal: Current Research in Pharmacology and Drug Discovery

Article Title: Long-term, stable, targeted biodelivery and efficacy of GDNF from encapsulated cells in the rat and Goettingen miniature pig brain

doi: 10.1016/j.crphar.2020.04.001

Figure Lengend Snippet: GDNF secretion from encapsulated cells implanted into intact versus 6-OHDALesioned.

Article Snippet: A commercially available GDNF ELISA kit (DuoSet® for human GDNF R&D Systems, Minneapolis, MN) was used to quantify GDNF device secretion prior to implantation and again following retrieval from the brain.

Techniques:

Journal: Current Research in Pharmacology and Drug Discovery

Article Title: Long-term, stable, targeted biodelivery and efficacy of GDNF from encapsulated cells in the rat and Goettingen miniature pig brain

doi: 10.1016/j.crphar.2020.04.001

Figure Lengend Snippet: Photomicrographs of substantia nigra stained for TH immunoreactivity illustrating loss of dopaminergic neurons following 6-OHDA and preservation of those neurons following GDNF treatment (a). Quantitation of dopaminergic neurons in the substantia nigra demonstrates virtually complete protection of lesioned neurons (b). Data are presented as mean ​± ​SEM % of TH-positive neurons relative to the intact, non-lesioned hemisphere of 8 animals per group. Scale bars, 200 ​μm. Multiple comparisons were based on significant main effects of treatment resulting from the ANOVA described in Results: ∗∗p<0.01.

Article Snippet: A commercially available GDNF ELISA kit (DuoSet® for human GDNF R&D Systems, Minneapolis, MN) was used to quantify GDNF device secretion prior to implantation and again following retrieval from the brain.

Techniques: Staining, Preserving, Quantitation Assay

Journal: Current Research in Pharmacology and Drug Discovery

Article Title: Long-term, stable, targeted biodelivery and efficacy of GDNF from encapsulated cells in the rat and Goettingen miniature pig brain

doi: 10.1016/j.crphar.2020.04.001

Figure Lengend Snippet: Photomicrographs of striatum stained for TH immunoreactivity (a) and quantitation of TH fiber density (b) following 6-OHDA with and without GDNF treatment illustrates a robust ability of GDNF to preserve dopaminergic fibers. Scale bars ​= ​500 ​μm Data are presented as mean ​± ​SEM % of TH-positive fibers relative to the intact, non-lesioned hemisphere of 8 animals per group. Multiple comparisons were based on significant main effects of treatment resulting from the ANOVA described in Results: ∗∗p<0.01.

Article Snippet: A commercially available GDNF ELISA kit (DuoSet® for human GDNF R&D Systems, Minneapolis, MN) was used to quantify GDNF device secretion prior to implantation and again following retrieval from the brain.

Techniques: Staining, Quantitation Assay

Journal: Current Research in Pharmacology and Drug Discovery

Article Title: Long-term, stable, targeted biodelivery and efficacy of GDNF from encapsulated cells in the rat and Goettingen miniature pig brain

doi: 10.1016/j.crphar.2020.04.001

Figure Lengend Snippet: Normal weight gain and neurological performance is maintained in lesioned animals by implantation of GDNF devices. (a) Body weights of animals with and without GDNF treatment. (b–d) Performance in the cylinder, placing, and stepping tests are all significantly impaired at 2 and 4 weeks following intrastriatal injections of 6-OHDA. In contrast, performance on each of these tests is preserved by GNDF implants with treated animals performing comparably to pre-surgery levels (pre-implantation and pre-lesion). Data are presented as mean ​± ​SEM of 8 animals per group. Multiple comparisons were based on significant main effects or interactions resulting from ANOVAs that are described in Results: ∗∗p<0.01.

Article Snippet: A commercially available GDNF ELISA kit (DuoSet® for human GDNF R&D Systems, Minneapolis, MN) was used to quantify GDNF device secretion prior to implantation and again following retrieval from the brain.

Techniques:

Journal: Current Research in Pharmacology and Drug Discovery

Article Title: Long-term, stable, targeted biodelivery and efficacy of GDNF from encapsulated cells in the rat and Goettingen miniature pig brain

doi: 10.1016/j.crphar.2020.04.001

Figure Lengend Snippet: Long-term, normal weight gain (a) and neurological performance in rats receiving GDNF implants 1 month post 6-OHDA. Then evaluated for 62 weeks. Testing on both the cylinder and placing tests (b-c) revealed a smooth and ever growing improvement in performance over the 62 week testing period. Data are presented as mean ​± ​SEM of 8 animals per group. Multiple comparisons were based on significant main effects or interactions resulting from ANOVAs that are described in Results: ∗p<0.05.

Article Snippet: A commercially available GDNF ELISA kit (DuoSet® for human GDNF R&D Systems, Minneapolis, MN) was used to quantify GDNF device secretion prior to implantation and again following retrieval from the brain.

Techniques:

Journal: Current Research in Pharmacology and Drug Discovery

Article Title: Long-term, stable, targeted biodelivery and efficacy of GDNF from encapsulated cells in the rat and Goettingen miniature pig brain

doi: 10.1016/j.crphar.2020.04.001

Figure Lengend Snippet: GDNF immunoreactivity in rat striatum 62 weeks following implantation of encapsulated GDNF-secreting cells (a) is associated with a pronounced preservation of TH-positive fibers that normally occurs post 6-OHDA (b–c). Scale bars ​= ​500 ​μm.

Article Snippet: A commercially available GDNF ELISA kit (DuoSet® for human GDNF R&D Systems, Minneapolis, MN) was used to quantify GDNF device secretion prior to implantation and again following retrieval from the brain.

Techniques: Preserving

Journal: Current Research in Pharmacology and Drug Discovery

Article Title: Long-term, stable, targeted biodelivery and efficacy of GDNF from encapsulated cells in the rat and Goettingen miniature pig brain

doi: 10.1016/j.crphar.2020.04.001

Figure Lengend Snippet: Implantation of 2 clinical-sized devices into the right putamen of minipigs delivers GDNF for 12 weeks and produces widespread delivery of GDNF (a–b) throughout the implanted striatum. GDNF delivery is also associated with a profound biological upregulation on dopaminergic function as determined by enhanced TH-immunoreactivity and quantitative optical densitometry in the striatum (c–d). Scale bars ​= ​5 ​mm. Data are presented as mean ​± ​SEM of 6 animals. ∗p<0.05; ∗∗p<0.01, Student's t test.

Article Snippet: A commercially available GDNF ELISA kit (DuoSet® for human GDNF R&D Systems, Minneapolis, MN) was used to quantify GDNF device secretion prior to implantation and again following retrieval from the brain.

Techniques:

Journal: Current Research in Pharmacology and Drug Discovery

Article Title: Long-term, stable, targeted biodelivery and efficacy of GDNF from encapsulated cells in the rat and Goettingen miniature pig brain

doi: 10.1016/j.crphar.2020.04.001

Figure Lengend Snippet: Serum and CSF measurements following GDNF implants in minipig Brain.

Article Snippet: A commercially available GDNF ELISA kit (DuoSet® for human GDNF R&D Systems, Minneapolis, MN) was used to quantify GDNF device secretion prior to implantation and again following retrieval from the brain.

Techniques:

Journal: Tissue Engineering Part A

Article Title: Finely Tuned Temporal and Spatial Delivery of GDNF Promotes Enhanced Nerve Regeneration in a Long Nerve Defect Model

doi: 10.1089/ten.tea.2015.0311

Figure Lengend Snippet: Figure 1: Representation of isograft and ANA surgical procedure. A) A representative schematic of an intermediate GDNF delivery time point in which GDNF is released by both a HBDS and Tet-on GDNF SCs under 6 weeks of doxycycline induction. B) The sciatic nerve is exposed and separated after the trifurcation to spare the sural nerve. The common peroneal and tibial nerves are transected approximately 5 mm distal to the trifurcation and a 3 cm reverse isograft or ANA is used to bridge the proximal and distal nerve stumps. 3-4 sutures were used to co-apt the grafts to the host tissue. For animals receiving SCs, SCs were injected beneath the epineurium distal to the graft area. Representative images are shown of the 3 cm GDNF-DS modified ANA prior to grafting and immediately after. C) tdTomato control vector and D) merged GFP and tdTomato expression is observed 2 weeks after injection into distal nerve stumps. Images are taken at 0.63x magnification. Red indicates control vector-labeled SCs, green represents Thy1-GFP axons.

Article Snippet: In vitro Bioactivity Assay GDNF expression from GDNF-SCs and tdTomato-SCs (in vivo control vector) was analyzed using a GDNF ELISA Duoset kit (R&D Systems) according to previous methods (31).

Techniques: Injection, Modification, Control, Plasmid Preparation, Expressing, Labeling

Journal: Tissue Engineering Part A

Article Title: Finely Tuned Temporal and Spatial Delivery of GDNF Promotes Enhanced Nerve Regeneration in a Long Nerve Defect Model

doi: 10.1089/ten.tea.2015.0311

Figure Lengend Snippet: Figure 2: SCs were successfully transduced with a Tet-on GDNF LV vector. A) A cartoon depiction of the Tet-on GDNF LV vector shows GDNF expression induced with tetracycline (or its analog doxycycline). Tetracycline binds to the reverse transcriptase activator (rtTA3) and then together the tetracycline and rtTA3 activate the tetracycline response element (TRE) driving GDNF and DsRed expression. B) Merged bright-field and fluorescent image is shown of Tet-on GDNF-SCs just prior to injection. C) GDNF released from control vector-SCs and GDNF-SCs after doxycycline induction. D) Average neurite length showed no difference between WT and control vector-SCs. Neurite length significantly increased in cultures with GDNF over- expressing SCs. Data is represented by the mean ± SEM. * denotes statistical significance from WT-SCs (p<0.05) E) mRNA was extracted from the distal nerve segments of GDNF-SC and control vector-SC transplanted animals and GDNF mRNA levels were examined. Compared to control vector-SC animals, GDNF mRNA was almost 20-fold higher in the distal nerve segment where SCs over-expressed GDNF. Dotted line marks up-regulation.

Article Snippet: In vitro Bioactivity Assay GDNF expression from GDNF-SCs and tdTomato-SCs (in vivo control vector) was analyzed using a GDNF ELISA Duoset kit (R&D Systems) according to previous methods (31).

Techniques: Transduction, Plasmid Preparation, Expressing, Reverse Transcription, Injection, Control

Journal: Tissue Engineering Part A

Article Title: Finely Tuned Temporal and Spatial Delivery of GDNF Promotes Enhanced Nerve Regeneration in a Long Nerve Defect Model

doi: 10.1089/ten.tea.2015.0311

Figure Lengend Snippet: Figure 3: Live imaging of GFP axons highlights differences in regeneration. A) Fluorescent images of Thy1-GFP axons at the end of the 8-week recovery period for the control and experimental conditions are shown. By the end of the recovery period, regeneration of GFP+ axons past the graft area can be observed clearly in the isograft control group. The GDNF-DS group also indicates distal growth of GFP+ axons, but regeneration appears greatly reduced in the ANA group and slightly reduced in the control vector-SC group. When doxycycline was removed too early at 4 weeks or continued for the entire 8 weeks (with or without the GDNF- DS), GFP+ axons become entrapped within the graft. Only 6 weeks of doxycycline shows GFP+ axons past the distal suture line. White arrows denote start and end of graft region. Scale bar = 3 mm. B) Average length of GFP+ axons over the 8 week recovery period is shown. Isografts had significantly greater length of GFP fluorescence over time. Only at 8 weeks does is stand out that

Article Snippet: In vitro Bioactivity Assay GDNF expression from GDNF-SCs and tdTomato-SCs (in vivo control vector) was analyzed using a GDNF ELISA Duoset kit (R&D Systems) according to previous methods (31).

Techniques: Imaging, Control, Plasmid Preparation, Fluorescence

Journal: Tissue Engineering Part A

Article Title: Finely Tuned Temporal and Spatial Delivery of GDNF Promotes Enhanced Nerve Regeneration in a Long Nerve Defect Model

doi: 10.1089/ten.tea.2015.0311

Figure Lengend Snippet: Figure 5: GDNF delivery from the DS-modified ANA and 6 weeks of GDNF over- expression improves total axon number and axon density. Total number of axons in the A) midgraft and B) distal nerve sections for each group was quantified. Axon density was quantified in the A) midgraft and B) distal nerve segments. Data is represented by the mean ± SEM (n≥ 6 animals per group). * denotes statistical significance from isograft (p<0.05), ^ significance from DS ANA (p<0.05), + significance from GDNF-SCs DS Dox6 (p<0.05).

Article Snippet: In vitro Bioactivity Assay GDNF expression from GDNF-SCs and tdTomato-SCs (in vivo control vector) was analyzed using a GDNF ELISA Duoset kit (R&D Systems) according to previous methods (31).

Techniques: Modification, Over Expression

Journal: Tissue Engineering Part A

Article Title: Finely Tuned Temporal and Spatial Delivery of GDNF Promotes Enhanced Nerve Regeneration in a Long Nerve Defect Model

doi: 10.1089/ten.tea.2015.0311

Figure Lengend Snippet: Figure 6: The quality of nerve regeneration was improved for DS-modified ANA and 6 weeks of GDNF over-expression. The percent neural tissue in the A) midgraft and B) distal nerve for each group was quantified. The percent myelin debris was quantified in the A) midgraft and B) distal nerve segments. Data is represented by the mean ± SEM (n≥ 6 animals per group). * denotes statistical significance from isograft (p<0.05), ^ significance from DS (p<0.05), + significance from GDNF-SCs DS Dox6 (p<0.05).

Article Snippet: In vitro Bioactivity Assay GDNF expression from GDNF-SCs and tdTomato-SCs (in vivo control vector) was analyzed using a GDNF ELISA Duoset kit (R&D Systems) according to previous methods (31).

Techniques: Modification, Over Expression

Journal: Tissue Engineering Part A

Article Title: Finely Tuned Temporal and Spatial Delivery of GDNF Promotes Enhanced Nerve Regeneration in a Long Nerve Defect Model

doi: 10.1089/ten.tea.2015.0311

Figure Lengend Snippet: Figure 7: The timing or method of GDNF delivery did not affect fiber width distribution. A) Distribution of the average fiber width of regenerated fibers in the A) midgraft and B) distal nerve was quantified. Data is represented by the mean ± SEM (n≥ 3 animals per group, animals with 0 regenerated fibers were not included).

Article Snippet: In vitro Bioactivity Assay GDNF expression from GDNF-SCs and tdTomato-SCs (in vivo control vector) was analyzed using a GDNF ELISA Duoset kit (R&D Systems) according to previous methods (31).

Techniques:

Journal: Tissue Engineering Part A

Article Title: Finely Tuned Temporal and Spatial Delivery of GDNF Promotes Enhanced Nerve Regeneration in a Long Nerve Defect Model

doi: 10.1089/ten.tea.2015.0311

Figure Lengend Snippet: Figure 8: Six weeks of GDNF over-expression from distally transplanted SCs leads to enhanced muscle mass recovery. A) Percent muscle mass recovery of the A) gastrocnemius muscle and B) tibialis anterior muscle compared to the unoperated control. Data is represented by the mean ± SEM (n≥ 6 animals per group). * denotes statistical significance from isograft (p<0.05), + significance from GDNF-SCs DS Dox6 (p<0.05), # denotes significance from all groups except isograft (p<0.05).

Article Snippet: In vitro Bioactivity Assay GDNF expression from GDNF-SCs and tdTomato-SCs (in vivo control vector) was analyzed using a GDNF ELISA Duoset kit (R&D Systems) according to previous methods (31).

Techniques: Over Expression, Control