primary human neonatal dermal fibroblasts (ndfs  (AllCells LLC)

Journal: Cell reports

Article Title: Pressure-Driven Mitochondrial Transfer Pipeline Generates Mammalian Cells of Desired Genetic Combinations and Fates

doi: 10.1016/j.celrep.2020.108562

Figure Lengend Snippet: (A) Selection workflow (in days) for generating SIMR cells from ρ0 primary human fibroblasts and SIMR clone generation efficiency data. Mitochondria from ~3 × 10 7 peripheral blood mononuclear cells (PBMC1) were MitoPunch transferred into BJ ρ0, NDF ρ0, or ADF ρ0 recipient fibroblasts. After selection, SIMR colonies were stained with crystal violet and quantified. Clone counts from a single representative mitochondrial transfer into ~1 × 10 5 recipient ρ0 fibroblasts are indicated. (B) D-loop hypervariable region mtDNA sequences from native BJ, PBMC1, and BJ ρ0+PBMC1 SIMR fibroblasts. Arrows denote single-nucleotide polymorphisms. (C) Major histocompatibility complex (MHC) class I HLA A, B, and C locus genotyping using OptiType v.1.3.1 for native BJ, BJ ρ0, and BJ ρ0+PBMC1 SIMR fibroblasts. (D) OCR measurements for ~1.5 3 10 4 native BJ, BJ ρ0, and BJ ρ0+PBMC1 SIMR fibroblasts by the Seahorse XF96 Extracellular Flux Analyzer. Values were calculated by standard procedures (see ). Data are the means ± SD of four technical replicates. Statistical significance by unpaired, two-tailed Student’s t test. *p ≨ 0.05; ***p ≨ 0.001 (E) Representative images of native BJ, BJ ρ0, and BJ ρ0+PBMC1 SIMR fibroblasts immunostained for double-stranded DNA (dsDNA) (green) and TOM20 (red) with colocalization indicated (yellow). Images (1003) were acquired on a Leica SP8 confocal microscope. Scale bars, 15 μm. See also and .

Article Snippet: Primary, non-transformed human fibroblast sources include BJ (ATCC, Cat. # CRL-2522), ADF (ATCC, Cat. # PCS-201–012), and NDF (ATCC, Cat. # PCS-201–010).

Techniques: Selection, Staining, Immunopeptidomics, Two Tailed Test, Microscopy