# Application of Conditioned Medium for In Vitro Modeling and Repair of Respiratory Tissue
## 3 Results
### Application of CM from Non-Respiratory Cells or Tissues
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**CELL TYPE** & \begin{tabular}{c} **CM CHARACTER-** \\ **ISTICS** \\ \end{tabular} & \begin{tabular}{c} **EXPERIMENTAL** \\ **EVALUATION** \\ \end{tabular} & \multicolumn{2}{c}{**RESULTS AND CONCLUSIONS**} \\ \hline  & & & - & MSC stimulated with TNF-a and IL-1ß \(\uparrow\) FGF2, HGF, HBEGF, IL-6 gene expression & - \\ Human & From MSC & & - & MSC-CM-STM \(\uparrow\) wound closure rate in NCI-H292. \\ pulmonary & stimulated with TNF-a and IL-1ß expression, & & - & MSC-CM \(\uparrow\) wound closure rate in ALI-PBEC culture compared to NCI-H292; MSC-CM-STM exerted the same effect as DMEM-STM in ALI-PBEC culture. \\ carcinoma cells (SCI-H292) and MSC only (MSC-CM) & ERK1/2 phosphorylation & - & MSC-CM-STM \(\uparrow\) ERK1/2 phosphorylation compared to control, which was mediated by the EGFR pathway. \\ MSC [57] & cultured for 24 h & & - & Inhibition of EGFR pathway \(\downarrow\) MSC-CM-STM effect on wound closure rate, proving EGFR was the predominant pathway. \\  & & & - & MSC are a viable option for cell-based therapy of inflammatory lung diseases such as COPD. \\ \hline  & & & - & ADSC-CM was protective against CSE and TGF-61-induced cell death and EMT, as evidenced by \(\uparrow\) cell viability and migration rates and \(\downarrow\) LDH levels. This effect was associated with the maintenance of E-cadherin levels, which inhibited EMT activation. \\ Lung carcinoma EC (A549), hubmen bronchiole cells (ADSC) cultured for 24 h & \begin{tabular}{c} Immunoblotting, \\ fluorescence \\ detection, cell \\ migration assay \\ \end{tabular} & - & \begin{tabular}{c} A549 in A549-CM and a-MEM had \(\downarrow\) viability compared to A549 alone; CS led to significant \(\downarrow\) in cell numbers and \(\uparrow\) LDH levels in A549. \\ ADSC-CM prevented CSE-induced EMT and reversed the progressive decrease of epithelial marker expression associated with TGF-1 therapy. \\ Stem cell-CM protected against cell death or CSE- and TGF-1-induced EMT, indicating its potential as a new target for lung cancer treatment. \\ \hline  & & & - & MSC-CM \(\uparrow\) Na\({}^{+}\) transport, maximal ENaC, maximal Na\({}^{+}\) and K-ATPase activity in FDLE monolayer. \\  & & - & MSC-CM \(\uparrow\) mRNA expression involved in Na\({}^{+}\) transport, proving the earlier \(\uparrow\) in Na\({}^{+}\) transport. \\ Fetal distal lung EC (FDLE) [59] & From umbilical cord mesenchymal stromal cells cultured for 72 h & \begin{tabular}{c} Immunofluorescence \\ staining, cell \\ permeability \\ assay, chamber \\ measurement, \\ gene expression \\ analyses, MTT \\ \end{tabular} & \begin{tabular}{c} - \\ staining, cell \\ permeability \\ assay, chamber \\ measurement, \\ gene expression \\ analyses, MTT \\ \end{tabular} & \begin{tabular}{c} _Strip_ and _Strip_ in MSC-CM induced, aiding functional maturation. \\ MSC-CM did not improve epithelial barrier function and proliferation. \\ ART and PI3K signaling contributed to MSC-CM-induced Na\({}^{+}\) transport and ENaC activity. \\ Rachl signaling could be involved in MSC-CM-stimulated ENaC activity. \\ MSC-CM significantly accelerated functional and structural maturation of fetal lungs, with effects mediated at least in part by the PI3K–AKT and Rac1 signaling pathways. \\ \hline  & & - & PMA-treated mCM \(\downarrow\) viability in A549 culture, triggered by inflammatory cytokines present in mCM, i.e., TNF-a and IL-8. \\  & & - & Aesculetin \(\uparrow\) viability in A549 + mCM compared to no aseculetin. \\  & & - & Aesculetin \(\uparrow\) E-cadherin in A549 with IL-8-induced mCM compared to no aseculetin, suppressing EMT induction on mCM. \\ Lung carcinoma EC (A549) [52] & \begin{tabular}{c} From THP-1 \\ monocyte-derived \\ macrophages (mCM) \\ cultured for 24 h \\ \end{tabular} & \
\begin{tabular}{c} ELISA, western \\ blotting, viability \\ assay, immunocytochemistry \\ \end{tabular} & - &
\begin{tabular}{c} Aesculetin \(\downarrow\) TIMP-1 and TIMP-2 expression in A549 with IL-8-induced mCM compared to no aseculetin. \\ ANCuletin \(\downarrow\) inflammatory macrophage infiltration in PHMG-induced mice compared to control. \\ Resculetin \(\downarrow\) CXCR2 induction in PHMG-induced mice. \\ Aesculetin exerted inhibitory effects on lung fibrosis and alveolar epithelial barrier breakdown caused by monocyte-derived macrophage infiltration. \\ \hline \hline \end{tabular}
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