Journal: Cell Reports Medicine

Article Title: Myeloid cells coordinately induce glioma cell-intrinsic and cell-extrinsic pathways for chemoresistance via GP130 signaling

doi: 10.1016/j.xcrm.2024.101658

Figure Lengend Snippet:

Article Snippet: Ciliary Neurotrophic Factor (CNTF) human , MedChemExpress , Cat#: HY-P7145.

Techniques: Plasmid Preparation, Recombinant, Transfection, Fluorescence, Staining, Reverse Transcription, Expressing, Liposomes, Mutagenesis, shRNA, Control, Construct, Software, Imaging, Functional Assay, Dissection, Sequencing, Real-time Polymerase Chain Reaction

Journal: EMBO Reports

Article Title: SERTM2: a neuroactive player in the world of micropeptides

doi: 10.1038/s44319-025-00404-w

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Recombinant Human CNTF , Peprotech , cat# AF-450-13.

Techniques: Modification, Sterility, Knock-Out, Recombinant, Transfection, Protease Inhibitor, Membrane, Staining, Electron Microscopy, Irradiation, Control, Sequencing, Cloning, Inverse PCR, Saline, Plasmid Preparation, DNA Extraction, Software, Microscopy, Subcloning

Journal: PLOS ONE

Article Title: Efficient derivation of functional astrocytes from human induced pluripotent stem cells (hiPSCs)

doi: 10.1371/journal.pone.0313514

Figure Lengend Snippet: Methods published for rapid (≤ 60 days) in vitro generation of astrocytes from stem cell-derived NPCs.

Article Snippet: On Day 21, astrocyte progenitor cells were plated into Matrigel-coated plates (25.000–75.000/cm 2 ) in serum-free astrocyte maturation medium (AMM), which was AGM supplemented with 1x AGS, 1% Pen/Strep and 20 ng/mL recombinant human ciliary neurotrophic factor (CNTF, provided by Peprotech).

Techniques: In Vitro, Glutathione Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Accelerated innervation of biofabricated skeletal muscle implants containing a neurotrophic factor delivery system

doi: 10.3389/fbioe.2024.1476370

Figure Lengend Snippet: Bioprinted skeletal muscle constructs. (A) A pair of fully printed skeletal muscle constructs with a neurotrophic factor delivery system. They have dimensions of 10 mm × 10 mm × 4 mm (W × L × H) and are composed of eight layers of cell-laden fibrinogen-based bioink with sacrificial bioink spacers. The cell-laden bioink contains CNTF/GDNF-loaded microspheres. In addition, an anchoring structure composed of PCL is deposited as a square frame at the periphery of the implant. (A′) Diagram of the microscale organization of the muscle construct. Created with BioRender. Abbreviations: achr –AChR cluster; f – fibrin hydrogel; m –muscle cell; ms –microsphere. (B) Differentiated muscle cells (red) with induced pre-formed AChR clusters (green) in the muscle construct prior to implantation. The sample was immunostained with antibodies against myosin heavy chain (MHC, red) and AChR (green). The image is a maximum intensity Z-projection of a confocal stack.

Article Snippet: Briefly, commercially available recombinant human CNTF (R&D, 257-NT) and GDNF (Millipore Sigma, GF322) were reconstituted as per the manufacturers’ instructions and combined at a concentration of 1 mg/mL in PBS containing 0.1% (w/v) bovine serum albumin (BSA).

Techniques: Construct

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Accelerated innervation of biofabricated skeletal muscle implants containing a neurotrophic factor delivery system

doi: 10.3389/fbioe.2024.1476370

Figure Lengend Snippet: Representative micrographs of chick embryo dorsal root ganglia (DRGs) cultured on 3D bioprinted skeletal muscle constructs in the in vitro neurite outgrowth assay. In the Control group (A–C) , the constructs contained no neurotrophic factors. The microsphere-free constructs in the NFs group (A′–C′) contained CNTF and GDNF directly mixed into the hydrogel of the construct. The constructs in the MSs group (A′′–C′′) contained the matching load of the neurotrophic factors encapsulated in PLGA microspheres. The ganglia were cultured on the muscle constructs for 2 days (A, A′′) , 7 days (B, B′′) , and 14 days (C, C′′) . Micrographs in the right column (1A′′–1C′′) are representative high-magnification images [of the boxed areas in (A′′–C′′) , respectively]. The samples were then fixed and immunostained with anti-neurofilament antibodies. All images are maximum intensity Z-projections of confocal stacks. The neurites were traced with the NeuronJ plugin in the Fiji/ImageJ software and are shown in magenta.

Article Snippet: Briefly, commercially available recombinant human CNTF (R&D, 257-NT) and GDNF (Millipore Sigma, GF322) were reconstituted as per the manufacturers’ instructions and combined at a concentration of 1 mg/mL in PBS containing 0.1% (w/v) bovine serum albumin (BSA).

Techniques: Cell Culture, Construct, In Vitro, Neurite Outgrowth Assay, Control, Software

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Accelerated innervation of biofabricated skeletal muscle implants containing a neurotrophic factor delivery system

doi: 10.3389/fbioe.2024.1476370

Figure Lengend Snippet: Representative micrographs of the histological organization of the tissue samples from the in vivo transposed nerve study at 4 weeks (A–A‴) , 8 weeks (B–B‴) , and 12 weeks (C–C‴) post-implantation. Hematoxylin and eosin staining. The Acellular cohort involves fibrin hydrogel implants that contain no cells and no neurotrophic factors. The Control group involves biofabricated muscle implants with 30 million hMPCs per mL of hydrogel, but no extrinsic neurotrophic factors. The third group (NFs ) contains CNTF and GDNF freely dissolved in the extracellular matrix of the implant. The fourth group (MSs) contains the matching load of the neurotrophic factors encapsulated in PLGA microspheres suspended throughout the hydrogel of the construct. The implanted constructs are outlined. Note that in the MSs cohort, the microspheres are abundant in the extracellular matrix of the implant at 4 weeks post-implantation. After 8 weeks, they become much smaller in size and are no longer detectable after 12 weeks. This indicates that the microspheres slowly dissolve in time and release the encapsulated CNTF and GDNF. cpn –transposed common peroneal nerve; ms –CNTF/GDNF-loaded PLGA microspheres.

Article Snippet: Briefly, commercially available recombinant human CNTF (R&D, 257-NT) and GDNF (Millipore Sigma, GF322) were reconstituted as per the manufacturers’ instructions and combined at a concentration of 1 mg/mL in PBS containing 0.1% (w/v) bovine serum albumin (BSA).

Techniques: In Vivo, Staining, Control, Construct

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Accelerated innervation of biofabricated skeletal muscle implants containing a neurotrophic factor delivery system

doi: 10.3389/fbioe.2024.1476370

Figure Lengend Snippet: Compound muscle action potential (CMAP) in the transposed nerve model study at 4, 8, and 12 weeks post-implantation. The Acellular cohort received bioprinted implants containing only fibrin hydrogel with no cells and no intrinsic neurotrophic factors. The Control group contains cellularized biofabricated muscle implants without any extrinsic neurotrophic factors. The third group (NFs) contains CNTF and GDNF freely dissolved in the extracellular matrix of the cellularized implant. The fourth group (MSs) contains the matching load of the neurotrophic factors encapsulated in PLGA microspheres suspended throughout the extracellular matrix of the construct. (A) CMAP amplitude. At 12 weeks post-implantation, the CMAP recorded from the constructs of the MSs cohort shows a trend to be higher than in the other two cohorts with cellularized constructs (the Control and NFs cohorts), and was the only one showing a statistically significant difference from the Acellular cohort. These data indicate that the CNTF and GDNF released from the PLGA microspheres facilitate innervation of the skeletal muscle implants after 12 weeks. This effect requires the microsphere-based neurotrophic factor delivery system and cannot be achieved by implementing the neurotrophic factors in the freely dissolved form. Three animals were analyzed in each treatment cohort (n = 3) * p < 0.05 (B) Representative waveforms.

Article Snippet: Briefly, commercially available recombinant human CNTF (R&D, 257-NT) and GDNF (Millipore Sigma, GF322) were reconstituted as per the manufacturers’ instructions and combined at a concentration of 1 mg/mL in PBS containing 0.1% (w/v) bovine serum albumin (BSA).

Techniques: Control, Construct

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Accelerated innervation of biofabricated skeletal muscle implants containing a neurotrophic factor delivery system

doi: 10.3389/fbioe.2024.1476370

Figure Lengend Snippet: Quantitative assessment of neurite sprouting from the transposed host common peroneal nerve in the implanted skeletal muscle constructs at 4, 8, and 12 weeks post-implantation. The Acellular cohort involves fibrin hydrogel implants with no cells and no extrinsic neurotrophic factors. The Contro l group contains cellularized biofabricated muscle implants without any extrinsic neurotrophic factors. The third group (NFs) contains CNTF and GDNF freely dissolved in the extracellular matrix of the implant. The fourth group (MSs) contains the matching load of the neurotrophic factors encapsulated in PLGA microspheres suspended throughout the hydrogel of the construct. There is no statistically significant difference among the treatment cohorts at 4 weeks post-implantation. At 8 weeks, the sprouting in the Control and MSs cohorts was higher than in the Acellular cohort. At 12 weeks, the constructs in the MSs cohort showed more extensive sprouting than any of the other three cohorts, indicating that only the CNTF/GDNF delivered in PLGA microspheres were capable of sustaining long-term neurite outgrowth. Three to four animals were analyzed in each treatment group at all three time points. * p < 0.05 , *** p < 0.001 .

Article Snippet: Briefly, commercially available recombinant human CNTF (R&D, 257-NT) and GDNF (Millipore Sigma, GF322) were reconstituted as per the manufacturers’ instructions and combined at a concentration of 1 mg/mL in PBS containing 0.1% (w/v) bovine serum albumin (BSA).

Techniques: Construct, Control