Ex4 (Anaspec, Fremont, CA) or IGF-1 (Shenandoah Biotechnology, Warwick, PA) was encapsulated into PLG microspheres using a double emulsion technique previously described (28 (link)). Two concentrations of PLG (3% or 6% w/w) and ratios of the lactide:glycolide polymers (50:50, 75:25 or a blend) were tested. Encapsulation efficiency was determined with 125I-labeled proteins.
Igf 1
Manufactured by Fujifilm
Sourced in United States
IGF-1 is a laboratory test kit designed to measure the levels of Insulin-like Growth Factor 1 (IGF-1) in biological samples. IGF-1 is a protein hormone that plays a key role in cell growth and development. The IGF-1 test kit provides a quantitative analysis of IGF-1 levels, which can be useful in the diagnosis and monitoring of various medical conditions.
Automatically generated - may contain errors
Lab products found in correlation
Epidermal growth factor (egf), by Fujifilm (2 mentions)
Neurobasal, by Thermo Fisher Scientific (2 mentions)
Dexamethasone (dex), by Fujifilm (2 mentions)
β glycerophosphate, by Fujifilm (2 mentions)
L ascorbic acid, by Fujifilm (2 mentions)
α mem, by Thermo Fisher Scientific (2 mentions)
Igf bp2, by Thermo Fisher Scientific (1 mentions)
2 3 dioxo 6 nitro 1 2 3 4 tetrahydrobenzo f quinoxaline 7 sulfonamide nbqx disodium salt, by Abcam (1 mentions)
Tgf β1, by Fujifilm (1 mentions)
Fibroblast growth factor 2 (fgf2), by Fujifilm (1 mentions)
α mem, by Fujifilm (1 mentions)
Mc3t3 e1 cells, by RIKEN BioResource Center (1 mentions)
7 protocols using igf 1
1
Microencapsulation of Protein Therapeutics
2
Optimizing Muscle Cell Differentiation
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To screen different muscle differentiation regimes, six different methods were tested: cessation of feeding (serum starvation); medium containing 2% FBS (reduced serum); a differentiation medium (reduced serum + additives) adapted from Messmer et al.3 (link)—reduced serum with insulin (Sigma Aldrich #I0516), 1-oleoyl lysophosphatidic acid (LPA) (Fisher Scientific #NC9401387), and transferrin (InVitria #777TRF029, Aurora, CO, USA); reduced serum medium with insulin-like growth factor 1 (IGF-1; Shenandoah Biotechnology #100-34, Warminster, PA, USA); reduced serum medium + additives with IGF-1; and a medium with an extracellular signal-regulated kinase inhibitor (ERKi, MilliPore Sigma #1049738-54-6, Burlington, MA, USA), adapted from Eigler et al.2 (link) More details on the media used are provided in Supplementary Table S1 .
Mack1 cells were seeded with laminin-511-E8 in experimental triplicate and, upon reaching 100% confluency, were fed with the different regimes listed above. After 10 days in culture, cells were fixed, stained with DAPI and MF-20, and images analyzed for the percent of MHC+ nuclei, as described earlier.
Mack1 cells were seeded with laminin-511-E8 in experimental triplicate and, upon reaching 100% confluency, were fed with the different regimes listed above. After 10 days in culture, cells were fixed, stained with DAPI and MF-20, and images analyzed for the percent of MHC+ nuclei, as described earlier.
3
Investigating IGF1 and IGFBP2 Signaling
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IGFBP2 was purchased from PeproTech, IGF1 was purchased from Shenandoah Biotechnology, the IGF1 antagonist JB1 was purchased from Sigma, and the AMPA/kainate receptor antagonist 2,3-Dioxo-6-nitro-1,2,3,4 tetrahydrobenzo[f]quinoxaline-7-sulfonamide (NBQX) disodium salt was purchased from Abcam. Doses for IGF1 and IGFBP2 were determined by dose response studies (Figure 1 ), and doses of JB1 and NBQX were chosen from Burgdorf et al. (2015a) . All drugs were administered in sterile saline vehicle at a volume of 1 mL/kg.
4
Serum-free Differentiation of Cells
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Throughout long-term culture, a population of cells at passages 2, 4, and 6 were cultured to confluency in serum-containing or serum-free conditions, and media was changed to a previously described serum-free differentiation media consisting of Neurobasal (Invitrogen #21103049, Carlsbad, CA, USA) and L15 (Invitrogen #11415064) basal media (1:1) supplemented with 1% antibiotic/antimycotic, 10 ng/mL insulin-like growth factor 1 (IGF-1; Shenandoah Biotechnology #100-34AF-100UG, Warminster, PA, USA) and 100 ng/mL epidermal growth factor (EGF; Shenandoah Biotechnology #100-26-500UG)10 (link). Cells were differentiated for 4-6 days (changing media every two days).
5
Neural Differentiation of Cells
6
Osteogenic Differentiation of Human iPSCs
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We previously reported an effective method for inducing osteogenesis in human iPSCs [21 (link),22 (link)]. Briefly, human iPSCs were incubated in the presence or absence of a SMO agonist or inhibitor in osteoblast differentiation medium (OBM). The OBM consisted of α-MEM (Invitrogen) supplemented with 10% FBS, 50 μg/ml L-ascorbic acid (Wako Pure Chemical Industries Ltd.), 10 mM β-glycerophosphate (Wako Pure Chemical Industries Ltd.), and 10 nM dexamethasone (Wako Pure Chemical Industries Ltd.). A combination of cytokines, referred to as FIT and comprising 25 ng/ml bFGF, 1 ng/ml TGF-β1 (Wako Pure Chemical Industries Ltd.), and 100 ng/ml IGF-1 (Wako Pure Chemical Industries Ltd.), was added on the following day (day 0). The iPSCs were cultured for an additional 10 days, with the OBM replenished every 3 days.
7
Osteoblast Differentiation of MC3T3-E1 Cells
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MC3T3-E1 cells were purchased from the RIKEN BioResource Center (Ibaraki, Japan). The collected cells were grown in Minimum Essential Medium Alpha (αMEM) (Thermo Fisher Scientific, Waltham, MA, USA) containing 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin. After trypsin treatment and rinsing with phosphate buffered saline (PBS), the MC3T3-E1 cells were resuspended in 50 μl medium at a seeding density of 1×10 6 cells. The cell solution was then seeded onto the ACS coated with 25 μl SPH from both sides. Next, the ACS samples were transferred into osteoblast differentiation medium (OBM) consisting of αMEM supplemented with 10% FBS, 50 μg/ml L-ascorbic acid (Wako Pure Chemical Industries Ltd.), 10 mM β-glycerophosphate (Wako Pure Chemical Industries Ltd.), and 10 nM dexamethasone (Wako Pure Chemical Industries Ltd.) with the following combinations of cytokines: 25 ng/ml FGF-2 (Wako Pure Chemical Industries Ltd.); 1 ng/ml TGF-b1 (Wako Pure Chemical Industries Ltd.); or 100 ng/ml IGF-1 (Wako Pure Chemical Industries Ltd.) 17, 31) . The samples were then cultured for 7 days at 37°C in a humidified atmosphere with 5% CO 2 . The medium was changed every 2 days.
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