A Microarray Analysis of the Effects of Serum-free Medium on Gene Expression Changes in Human Mesenchymal Stem Cells during the in Vitro Culture

AbstractBackgroundMesenchymal stem cells (MSCs) provide potential treatments for peritoneal fibrosis. However, MSCs cultured in media containing serum bring risks of infection and other problems. In this study, we compared the effect of human MSCs in serum-free medium (SF-MSCs) on peritoneal fibrosis with that of MSCs cultured in medium containing 10% fetal bovine serum (10%MSCs).MethodsPeritoneal fibrosis was induced by intraperitoneally injecting 0.1% chlorhexidine gluconate (CG). SF-MSCs or 10%MSCs were intraperitoneally administered 30 min after the CG injection. Ten days after the CG and MSC injections, we performed histological analyses and peritoneal equilibrium testing. In the in vitro experiments, we used transforming growth factor (TGF)-β1-stimulated human peritoneal mesothelial cells incubated in conditioned medium from MSCs to examine whether the SF-MSCs showed enhanced ability to produce antifibrotic humoral factors.ResultsHistological staining showed that the SF-MSCs significantly suppressed CG-induced cell accumulation and thickening compared with that of the 10%MSCs. Additionally, the SF-MSCs significantly inhibited mesenchymal cell expression, extracellular matrix protein deposition and inflammatory cell infiltration. Peritoneal equilibration testing showed that compared with administering 10%MSCs, administering SF-MSCs significantly reduced the functional impairments of the peritoneal membrane. The in vitro experiments showed that although the conditioned medium from MSCs suppressed TGF-β1 signaling, the suppression did not significantly differ between the SF-MSCs and 10%MSCs.ConclusionsSerum-free culture conditions can enhance the antifibrotic abilities of MSCs by suppressing inflammation. Administering ex vivo expanded SF-MSCs may be a potential therapy for preventing peritoneal fibrotic progression.

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Serum-Free Medium Enhances the Immunosuppressive and Antifibrotic Abilities of Mesenchymal Stem Cells Utilized in Experimental Renal Fibrosis

Stem Cells Translational Medicine ◽

10.1002/sctm.17-0284 ◽

2018 ◽

Vol 7 (12) ◽

pp. 893-905 ◽

Cited By ~ 14

Author(s):

Ken Yoshida ◽

Ayumu Nakashima ◽

Shigehiro Doi ◽

Toshinori Ueno ◽

Tomoe Okubo ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Renal Fibrosis ◽

Serum Free Medium ◽

Free Medium ◽

Serum Free

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Human Umbilical Cord-Derived Mesenchymal Stem Cells Do Not Undergo Malignant Transformation during Long-Term Culturing in Serum-Free Medium

PLoS ONE ◽

10.1371/journal.pone.0098565 ◽

2014 ◽

Vol 9 (6) ◽

pp. e98565 ◽

Cited By ~ 32

Author(s):

Gecai Chen ◽

Aihuan Yue ◽

Zhongbao Ruan ◽

Yigang Yin ◽

RuZhu Wang ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Malignant Transformation ◽

Umbilical Cord ◽

Human Umbilical Cord ◽

Serum Free Medium ◽

Free Medium ◽

Serum Free

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The Phosphate Source Influences Gene Expression and Quality of Mineralization during In Vitro Osteogenic Differentiation of Human Mesenchymal Stem Cells

PLoS ONE ◽

10.1371/journal.pone.0065943 ◽

2013 ◽

Vol 8 (6) ◽

pp. e65943 ◽

Cited By ~ 34

Author(s):

Luisa M. Schäck ◽

Sandra Noack ◽

Ramona Winkler ◽

Gesa Wißmann ◽

Peter Behrens ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Osteogenic Differentiation ◽

Human Mesenchymal Stem Cells ◽

Phosphate Source

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Comparative Study of the Biological Characteristics of Serum-Free and Fetal Bovine Serum-Contained Medium Cultured Umbilical Cord-Derived Mesenchymal Stem Cells

Blood ◽

10.1182/blood.v120.21.4737.4737 ◽

2012 ◽

Vol 120 (21) ◽

pp. 4737-4737 ◽

Cited By ~ 1

Author(s):

Guanghua Chen ◽

Ting Yang ◽

Man Qiao ◽

Huiwen Liu ◽

Wu Depei

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

T Cell ◽

Umbilical Cord ◽

Complete Medium ◽

Human Umbilical Cord ◽

Serum Free Medium ◽

Free Medium ◽

Early Passage ◽

Serum Free

Abstract Abstract 4737 Objective: To compare the difference of biological characteristics between human umbilical cord-derived mesenchymal stem cells (UC-MSC) cultured by serum free medium and fetal bovine serum-contained complete medium and to create a xenogeneic protein-free UC-MSC culture system. Methods: Healthy human umbilical cord segments were digested with collagenase. Umbilical cord-derived mesenchymal stem cells were cultured by serum free MesenCult-XF medium and FBS-based αMEM complete medium. We analysed the morphology, immunophenotype, expansion potential, trilineage differentiation potential, karyotype and immunosuppression of early passage of UC-MSC. Results: The average cell diameters of UC-MSC in suspension cultured by serum free medium and FBS-based medium are 26 (18–39) μm and 35 (20–61) μm, respectively. Cell expansion folds with serum free medium and FBS-based medium were (5.2±0.2) and (3.5±0.1) in the first five passage, respectively. The expansion potential of MSCs was significantly higher with serum free medium compared to FBS-based medium (P<0.05). A panel of markers as CD29, CD44, CD90, CD73, CD105 and HLA-ABC were expressed by human UC-MSC. Hematopoietic lineage markers CD34, CD45 and HLA-DR were not detectable on UC-MSC. The cpm were (4.57±0.14)×104, (2.04±0.16)×104 and(0.42±0.04)×104 when serum free medium cultured MSCs were added to the cultures at ratios MSCs/T cell of 1:100, 1:10 and 1:5. While the cpm were (4.57±0.14)×104, (2.04±0.16)×104 and(0.42±0.04)×104when serum free medium cultured UC-MSCs were added to the cultures. The immunosuppressive potential of serum free medium-cultured UC-MSC was higher than that of serum-contained medium cultured UC-MSC at three different ratios MSC/T cell (P<0.05). Conclusion Compare with serum-contained medium cultured early passage of UC-MSC, the cell diameter of serum free medium cultured MSCs was smaller and the expansion potential was higher. No xenogeneic proteins were presented in UC-MSC preparation when UC-MSC was cultured with serum free medium. Human UC-MSC suppresses T-cell proliferation in a dose-dependent manner. The immunosuppressive potential of UC-MSC was higher when cultured in serum free medium compared with FBS-based medium. Disclosures: No relevant conflicts of interest to declare.

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