culture conditions primary huvec  (PromoCell)

Journal: Bone

Article Title: Immature myeloid cells are critical for enhancing bone fracture healing through angiogenic cascade

doi: 10.1016/j.bone.2016.09.018

Figure Lengend Snippet: A) Gemcitabine treatment results in significant depletion of Gr-1+CD11b+ IMC. B) H & E staining and radiography of unhealed area in femoral segmental defect in wild-type mice (top) and gemcitabine treated mice (bottom) at 21 days post fracture. Relative quantitation of unhealed area following gemcitabine treatment, as compared to control is given on the right. Black arrows indicate fracture site, and failure to heal in gemcitabine-treated mice. (P < 0.05) C) Gemcitabine does not inhibit MSC differentiation or HUVEC tube formation: MSC were differentiated in osteogenic differentiation media in the presence of gemcitabine. After 14 days MSC were stained with Alizarin Red for calcium deposition (top panel). HUVEC cells were cultured on reduced growth factor matrigel with normal basal endothelial cell medium with or without gemcitabine (bottom panel). Quantitative analyses from replicate experiments are provided on the right for each panel for both osteoblast differentiation of MSC and HUVEC tube formation.

Article Snippet: Cells and culture conditions Human umbilical vein endothelial cells (HUVEC) were purchased from ATCC and grown in endothelial cell growth media (EGM -2, Lonza).

Techniques: Staining, Quantitation Assay, Control, Cell Culture

Journal: Bone

Article Title: Immature myeloid cells are critical for enhancing bone fracture healing through angiogenic cascade

doi: 10.1016/j.bone.2016.09.018

Figure Lengend Snippet: A) IMC isolated from naïve mice or mice after segmental fractures on days 1, 3, and 7were co-cultured with HUVEC in serum-free medium. IMC were placed in the bottom chamber and HUVEC in the top of the Boyden chamber. HUVEC cells capable of passing though pores were fixed and stained with neutral red after 4 hours. B) HUVEC cell motility was performed using scratch assay in dual chamber tissue culture plate. Upon reaching confluence, a linear defect was introduced to HUVEC monolayers with a 200 μl pipet tip. Naïve or activated IMC were placed in the upper compartment of Boyden chamber in serum-free media. Quantitative analysis of migrated HUVEC into the scratch area, provided on the right side, is expressed as percentage of migration as determined by closure of the gap width in indicated groups. (P<0.05 compared to control and the group co-cultured with naïve IMC). Quantitative analyses of migration, and motility data from triplicate experiments are provided on the right (*P<0.05;**P<0.01 compared to HUVEC alone, and using control IMC).

Article Snippet: Cells and culture conditions Human umbilical vein endothelial cells (HUVEC) were purchased from ATCC and grown in endothelial cell growth media (EGM -2, Lonza).

Techniques: Isolation, Cell Culture, Staining, Wound Healing Assay, Migration, Control

Journal: Bone

Article Title: Immature myeloid cells are critical for enhancing bone fracture healing through angiogenic cascade

doi: 10.1016/j.bone.2016.09.018

Figure Lengend Snippet: A) IMC isolated from naïve or 1, 3, or 7 days after segmental defect are co-cultured directly with HUVEC. IMC bind directly to HUVEC. IMC are labeled with CFSE, HUVEC with PKH26, 20x magnification. B) Representative images of HUVEC and IMC co-culture on tube formation assay. C) Quantitative analysis of tube formation using Angiogenesis Analyzer in ImageJ, n = 3 and three 10x images per well (*P<0.05).

Article Snippet: Cells and culture conditions Human umbilical vein endothelial cells (HUVEC) were purchased from ATCC and grown in endothelial cell growth media (EGM -2, Lonza).

Techniques: Isolation, Cell Culture, Labeling, Co-Culture Assay, Tube Formation Assay