lpc medium  (MedChemExpress)

Journal: bioRxiv

Article Title: Respiratory Airway Secretory Cells act as Immune Sentinels in Human Distal Airways

doi: 10.1101/2025.03.24.644887

Figure Lengend Snippet: a , Schematic of proximal-distal patterning in developing human lung with representative lineage markers. b , Differentiation protocol workflow from human pluripotent stem cells (hPSCs) to distal lung organoids (d-LOs) via anterior foregut endoderm (AFE) spheroids dissociation. c , Immunofluorescence staining of NKX2-1, SOX2 and SOX9 in proximal (p-LO) and distal (d-LO) lung organoids at day 8. White arrows denote SOX2 bright NKX2-1 dim cells; arrowheads indicate SOX9 bright NKX2-1 bright cells. Scale bars, 20 μm. Representative images from three biologically independent experiments. d , Quantitative PCR analysis of NKX2-1 , SOX9 and SOX2 expression in AFE spheroids, d-LOs and p-LOs. Data presented as mean ± s.e.m. (n=3 biological replicates). P -values were calculated using two-tailed Student’s t -test with Welch’s correction. e , UMAP with annotations of different cell types from epithelial cell subclusters. f , Individual conditions of epithelial subclusters shown in ( e ). g , Proportional distribution of cell types between d-LO and p-LO. h , Violin plots showing expression levels of proximal and distal markers in p-LPC and d-LPC cell populations from d-LO and p-LO.

Article Snippet: Detached anterior foregut spheroids (days 5–7) were split into two groups: p-LO: Spheroids were embedded in growth factor-reduced Matrigel (Corning, 354230) and cultured in LPC medium (20 ng/ml BMP4 [MCE, HY-P7007], 10 ng/ml FGF7/KGF [MCE, HY-P70597], 10 ng/ml FGF10 [MCE, HY-P70695], 3 μM CHIR99021, 20 μM DAPT [MCE, HY-13027], and 50 nM retinoic acid [Sigma, R2625]). d-LO: Spheroids were washed with PBS, dissociated into single cells with Accutase, pelleted (300 × g, 5 min), and resuspended in growth factor-reduced Matrigel.

Techniques: Immunofluorescence, Staining, Real-time Polymerase Chain Reaction, Expressing, Two Tailed Test

Journal: bioRxiv

Article Title: Respiratory Airway Secretory Cells act as Immune Sentinels in Human Distal Airways

doi: 10.1101/2025.03.24.644887

Figure Lengend Snippet: a , Differentiation protocol workflow for generating proximal lung organoids (p-LOs) from hPSCs. b , Bright-field images comparing organoid morphology under untreated versus dissociation conditions during lung progenitor cell (LPC) induction. Scale bars, 500 μm (far left and middle two panels), 200 μm (right panels). Representative of three biologically independent experiments. c , Phase-contrast images of day 8 p-LOs and d-LOs counterstained with Hoechst. Scale bars, 500 μm. d , Organoid yield quantification (number per Matrigel droplet). e , Organoid diameter measurements. f , Representative flow cytometry analysis of NKX2-1 expression in p-LOs versus d-LOs. n = 3 independent experiments. g , h , Percentage ( g ) and mean fluorescence intensity (MFI) ( h ) of NKX2-1 + cells. i , qPCR analysis of lung-specific and endodermal lineage markers in p-LOs and d-LOs derived from hESC lines (H1, H9) and induced pluripotent stem cells (hiPSCs). d , e , g , h , i , Data are presented as mean ± s.e.m. (n=3 biological replicates). P -values calculated using two-tailed Student’s t -test.

Article Snippet: Detached anterior foregut spheroids (days 5–7) were split into two groups: p-LO: Spheroids were embedded in growth factor-reduced Matrigel (Corning, 354230) and cultured in LPC medium (20 ng/ml BMP4 [MCE, HY-P7007], 10 ng/ml FGF7/KGF [MCE, HY-P70597], 10 ng/ml FGF10 [MCE, HY-P70695], 3 μM CHIR99021, 20 μM DAPT [MCE, HY-13027], and 50 nM retinoic acid [Sigma, R2625]). d-LO: Spheroids were washed with PBS, dissociated into single cells with Accutase, pelleted (300 × g, 5 min), and resuspended in growth factor-reduced Matrigel.

Techniques: Flow Cytometry, Expressing, Fluorescence, Derivative Assay, Two Tailed Test