primary hsaepcs  (PromoCell)

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: BPIFA1 is a secreted biomarker of differentiating human airway epithelium

doi: 10.3389/fcimb.2022.1035566

Figure Lengend Snippet: Study Design and Transcriptomic Analysis. (A) Diagram of healthy human small airway epithelial cells (hSAEpCs) (donor #1) guided through differentiation in vitro on transwell insert (TWI) porous membranes. The arrows indicate days following air-liquid interface (ALI) that cells were lysed in situ and used for transcriptomic analysis. (B) K -means clustering of genes differentially expressed during airway epithelial cell differentiation in vitro (these are genes with fold change [FC] > 8 and p < 0.01). D: days post-ALI; n = 3 replicates – i.e., TWI samples, per time point.

Article Snippet: Primary human small airway and bronchial epithelial cells were either purchased from commercial sources (Promocell, Lonza, LifeLine Cell Technology) or obtained from Drs. Fernando Holguin and Oliver Eickelberg.

Techniques: In Vitro, In Situ, Cell Differentiation

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: BPIFA1 is a secreted biomarker of differentiating human airway epithelium

doi: 10.3389/fcimb.2022.1035566

Figure Lengend Snippet: Transcriptomic Signature of In Vitro Differentiating Primary Human Airway Epithelial Cells. (A) Time course profile of expression of select genes from clusters plotted in Figure 1B . Left panel: microarray gene expression; right panel: hemi-nested qPCR validation of microarray data for days 0, 2, 8 and 28 post-ALI (same hSAEpC donor [donor #1]). (B) qPCR plots illustrating comparable patterns in expression of OMG, KRT14, STC1, BPIFA1, PLA2G7, TXNIP and S100A7 in three additional healthy human airway epithelial cell (hAEpC) donors, consistent with (A) . Top left (donor #2) and right (donor #3) panels: human bronchial epithelial cells [hBEpCs]: bottom left panel: hSAEpCs (donor #4). (C) Heatmap illustration of change in expression of genes in (A) and (B) across all tested donors (donors #1–4). OMG, oligodendrocyte myelin glycoprotein; KRT14, keratin 14; STC1, stanniocalcin 1; BPIFA1, BPI fold containing family A member 1; PLA2G7, phospholipase A2 group VII; TXNIP, thioredoxin interacting protein; S100A7, S100 calcium binding protein A7.

Article Snippet: Primary human small airway and bronchial epithelial cells were either purchased from commercial sources (Promocell, Lonza, LifeLine Cell Technology) or obtained from Drs. Fernando Holguin and Oliver Eickelberg.

Techniques: In Vitro, Expressing, Microarray, Binding Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: BPIFA1 is a secreted biomarker of differentiating human airway epithelium

doi: 10.3389/fcimb.2022.1035566

Figure Lengend Snippet: BPIFA1 Is a Secretory Biomarker of Differentiating Human Airway Epithelial Cells. Confirmation of BPIFA1 expression at protein level (secreted basolaterally into culture media) from six hAEpC donors (bronchial epithelial cells derived from two different donors [designated as yellow (donor #5) and red (donor #6)], and small airway epithelial cells derived from four separate donors [designated as purple (donor #7), orange (donor #8), blue (donor #9), and green (donor #10)]) with n = 4-7 replicates per donor. Data were analyzed by non-parametric Mann-Whitney test. *** p < 0.001, **** p < 0.0001, n.s., not significant.

Article Snippet: Primary human small airway and bronchial epithelial cells were either purchased from commercial sources (Promocell, Lonza, LifeLine Cell Technology) or obtained from Drs. Fernando Holguin and Oliver Eickelberg.

Techniques: Biomarker Assay, Expressing, Derivative Assay, MANN-WHITNEY

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: BPIFA1 is a secreted biomarker of differentiating human airway epithelium

doi: 10.3389/fcimb.2022.1035566

Figure Lengend Snippet: Change in Frequency of Airway Epithelial Sub-types and Secreted Inflammatory Proteins During In Vitro Differentiation. (A) The abundance of basal, goblet, Club, and ciliated cells were analyzed by flow cytometry on D0, 2, 8 and 35 post-ALI from four hAEpC donors (bronchial epithelial cells derived from two different donors [designated as yellow (donor #5) and red (donor #6)], and small airway epithelial cells derived from two separate donors [designated as blue (donor #9) and green (donor # 10)]) with n = 4 replicates per donor. Data were analyzed by non-parametric Mann-Whitney test. (B) Time course secretion analysis of inflammatory cytokines/chemokines for a representative human bronchial epithelial cell (donor #11) with n = 3–6 replicates. Data were analyzed by paired Student’s t -test test. * p < 0.05, ** p < 0.01, **** p < 0.0001. CSF2, colony-stimulating factor 2; IP10, interferon-gamma-inducible protein 10; MCP1, monocyte chemoattractant protein-1; RANTES, regulated upon activation; normal T cell expressed and presumably secreted; IL6, interleukin 6; IL8, interleukin 8.

Article Snippet: Primary human small airway and bronchial epithelial cells were either purchased from commercial sources (Promocell, Lonza, LifeLine Cell Technology) or obtained from Drs. Fernando Holguin and Oliver Eickelberg.

Techniques: In Vitro, Flow Cytometry, Derivative Assay, MANN-WHITNEY, Activation Assay