# Conditioned medium from human adipose-derived mesenchymal stromal cells can modulate cell migration and morphology of keratinocytes in vitro

**Authors:** Cristiano Rodrigues, Thaís Casagrande Paim, Carla Zanatelli, Elisa Vasconcellos Soares Prignon, Jéssica Gonçalves Azevedo, Liliana Ivet Sous Naasani, Márcia Rosângela Wink

PMC · DOI: 10.1007/s13577-026-01353-9 · Human Cell · 2026-02-03

## TL;DR

Human fat-derived stem cells release factors that change skin cell movement and shape, which could help in wound healing and skin regeneration.

## Contribution

The study shows that MSC secreted factors modulate keratinocyte migration and morphology via TGF-β1 signaling.

## Key findings

- MSC conditioned medium increased keratinocyte proliferation and migration.
- MSC secretome caused morphological changes like actin stress fibers and nuclei irregularities.
- TGF-β1 inhibition reduced keratinocyte migration by 76%.

## Abstract

The regenerative ability of the skin is orchestrated by different types of cells, including mesenchymal stromal cells (MSCs). Their role in wound healing is being widely studied due to their capacity to produce a secretome able to modulate their microenvironment. In this context, MSCs factors can be isolated using in vitro cell culture and be experimentally tested for a series of clinical conditions, such as skin repair. In this study, we produced conditioned medium (MSC-CM) using primary cultures from human adipose-derived MSCs. This medium was used as treatment in a culture of keratinocytes cell line from adult human skin (HaCaT) to investigate the modulation of their dynamics. We analyzed cell proliferation, migration, and changes on cell morphology by labeling the actin filaments and nuclei. The factors released by MSCs were able to improve both the proliferation and migration of keratinocytes. In addition, there was an increase in the amount of actin stress fibers, filopodia protrusions, and nuclei irregularities. The MSCs secretome modified the migratory patterns of keratinocytes, being observable through their morphological changes. At least in part, this modulation was caused by the TGF-β1 signaling, considering that its antagonist, SB 431542, lead to a reduction of approximately 76% in the migration of HaCaT cells through the porous membranes of transwell chambers. Together, our data contribute to a better understanding of the role of MSCs on keratinocytes during wound healing and reinforce the importance to investigate their potential in dermal regeneration therapies.

The online version contains supplementary material available at 10.1007/s13577-026-01353-9.

## Linked entities

- **Proteins:** TGFB1 (transforming growth factor beta 1)
- **Chemicals:** SB 431542 (PubChem CID 4521392)
- **Species:** Homo sapiens (taxon 9606)

## Full-text entities

- **Genes:** SMAD4 (SMAD family member 4) [NCBI Gene 4089] {aka DPC4, JIP, MADH4, MYHRS}, SMAD2 (SMAD family member 2) [NCBI Gene 4087] {aka CHTD8, JV18, JV18-1, LDS6, MADH2, MADR2}, INHBE (inhibin subunit beta E) [NCBI Gene 83729], ACVR1C (activin A receptor type 1C) [NCBI Gene 130399] {aka ACVRLK7, ALK7}, EGF (epidermal growth factor) [NCBI Gene 1950] {aka HOMG4, URG}, BMP1 (bone morphogenetic protein 1) [NCBI Gene 649] {aka OI13, PCOLC, PCP, TLD}, TGFBR1 (transforming growth factor beta receptor 1) [NCBI Gene 7046] {aka AAT5, ACVRLK4, ALK-5, ALK5, ESS1, LDS1}, IL6 (interleukin 6) [NCBI Gene 3569] {aka BSF-2, BSF2, CDF, HGF, HSF, IFN-beta-2}, CD14 (CD14 molecule) [NCBI Gene 929], TGFB1 (transforming growth factor beta 1) [NCBI Gene 7040] {aka CAEND1, CED, DPD1, IBDIMDE, LAP, TGF-beta1}, PTPRC (protein tyrosine phosphatase receptor type C) [NCBI Gene 5788] {aka B220, CD45, CD45R, GP180, IMD105, L-CA}, ACVR1B (activin A receptor type 1B) [NCBI Gene 91] {aka ACTRIB, ACVRLK4, ALK4, SKR2}, LAP (Laryngeal adductor paralysis) [NCBI Gene 7939], SMAD3 (SMAD family member 3) [NCBI Gene 4088] {aka HSPC193, HsT17436, JV15-2, LDS1C, LDS3, MADH3}, CD44 (CD44 molecule (IN blood group)) [NCBI Gene 960] {aka CDW44, CSPG8, ECM-III, ECMR-III, H-CAM, HCELL}, CD34 (CD34 molecule) [NCBI Gene 947], CXCL8 (C-X-C motif chemokine ligand 8) [NCBI Gene 3576] {aka GCP-1, GCP1, IL8, LECT, LUCT, LYNAP}, THY1 (Thy-1 cell surface antigen) [NCBI Gene 7070] {aka CD90, CDw90}, VEGFA (vascular endothelial growth factor A) [NCBI Gene 7422] {aka L-VEGF, MVCD1, VEGF, VPF}, ALK (ALK receptor tyrosine kinase) [NCBI Gene 238] {aka ALK1, CD246, NBLST3}
- **Diseases:** skin injury (MESH:D000069836), fibrosis (MESH:D005355), hypoxic (MESH:D002534), tumorigenic (MESH:D002471), trauma (MESH:D014947), burns (MESH:D002056)
- **Chemicals:** lipid (MESH:D008055), calcium (MESH:D002118), Alcian blue (MESH:D000423), DAPI (MESH:C007293), calcium phosphate (MESH:C020243), DMEM (-), Mit C (MESH:D016685), biotin (MESH:D001710), EDTA (MESH:D004492), SB 431542 (MESH:C459179), oxygen (MESH:D010100), PBS (MESH:D007854), CO2 (MESH:D002245), crystal violet (MESH:D005840), Oil Red O (MESH:C011049), trypan blue (MESH:D014343), Alizarin Red S (MESH:C004468)
- **Species:** Mus musculus (house mouse, species) [taxon 10090], Homo sapiens (human, species) [taxon 9606]
- **Cell lines:** B16 melanoma — Mus musculus (Mouse), Mouse melanoma, Cancer cell line (CVCL_F936), HaCaT — Homo sapiens (Human), Spontaneously immortalized cell line (CVCL_0038)

## Full text

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## Figures

7 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12864314/full.md

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Source: https://tomesphere.com/paper/PMC12864314