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Peripheral blood derived mononuclear cells enhance the migration and chondrogenic differentiation of multipotent mesenchymal stromal cells.

Hopper N, Wardale J, Howard D, Brooks R, Rushton N, Henson F - Stem Cells Int (2015)

Bottom Line: In scratch assay the wound closure was 55% higher after 3 hours in the PBMC stimulated test group (P = 0.002), migration rate was 9 times faster (P = 0.008), and total MSC migration was 25 times higher after 24 hours (P = 0.014).Analysis of MSCs by PCR array demonstrated that PBMCs induced the upregulation of genes associated with chondrogenic differentiation over 15-fold.In conclusion, PBMCs increase both MSC migration and differentiation suggesting that they are an ideal candidate for inclusion in regenerative medicine therapies aimed at cartilage repair.

View Article: PubMed Central - PubMed

Affiliation: Division of Trauma and Orthopaedic Surgery, University of Cambridge, Addenbrooke's Hospital, Hills Road, Cambridge BC2 0QQ, UK.

ABSTRACT
A major challenge in cartilage repair is the lack of chondrogenic cells migrating from healthy tissue into damaged areas and strategies to promote this should be developed. The aim of this study was to evaluate the effect of peripheral blood derived mononuclear cell (PBMC) stimulation on mesenchymal stromal cells (MSCs) derived from the infrapatellar fat pad of human OA knee. Cell migration was measured using an xCELLigence electronic migration chamber system in combination with scratch assays. Gene expression was quantified with stem cell PCR arrays and validated using quantitative real-time PCR (rtPCR). In both migration assays PBMCs increased MSC migration by comparison to control. In scratch assay the wound closure was 55% higher after 3 hours in the PBMC stimulated test group (P = 0.002), migration rate was 9 times faster (P = 0.008), and total MSC migration was 25 times higher after 24 hours (P = 0.014). Analysis of MSCs by PCR array demonstrated that PBMCs induced the upregulation of genes associated with chondrogenic differentiation over 15-fold. In conclusion, PBMCs increase both MSC migration and differentiation suggesting that they are an ideal candidate for inclusion in regenerative medicine therapies aimed at cartilage repair.

No MeSH data available.


Related in: MedlinePlus

Cell migration experiment of MSCs measured over 24 hours comparing the effect of PBMC stimulus (n = 4). (a) Wound closure rate measured at 3-hour time point (n = 4) in the scratch assay with direct cell-to-cell contact. (b) The total distance of the wound measured at 12-, 15-, and 24-hour time points is not to scale (n = 4, P < 0.0001) in the scratch assay. (c) The initial cell migration rate at 3-hour time point quantified real time with the xCELLigence system and (d) the total amount of cells migrated at the 24-hour time point quantified with cell index value by xCELLigence.
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fig2: Cell migration experiment of MSCs measured over 24 hours comparing the effect of PBMC stimulus (n = 4). (a) Wound closure rate measured at 3-hour time point (n = 4) in the scratch assay with direct cell-to-cell contact. (b) The total distance of the wound measured at 12-, 15-, and 24-hour time points is not to scale (n = 4, P < 0.0001) in the scratch assay. (c) The initial cell migration rate at 3-hour time point quantified real time with the xCELLigence system and (d) the total amount of cells migrated at the 24-hour time point quantified with cell index value by xCELLigence.

Mentions: Scratch assay showed that direct cell-to-cell PBMC stimulation increased the wound closure rate by 55.2% at 3 hours, P = 0.002 (Figure 2(a)). The distance of the gap was measured at 12-, 15-, and 24-hour time points (Figure 2(b)) and every time point the PBMC treated wound had smaller scratch area (P < 0.0001). To confirm the findings from the scratch assay and to assess if direct cell-to-cell contact is needed for the chemotactic effect, a second assay based on the Boyden chamber model (xCELLigence) was used to quantify cell migration. The real-time cell migration assay demonstrated that during the first 3 hours the MSC migration was 9 times faster in the PBMC stimulated test group (P = 0.008, Figure 2(c)). The results of the 24-hour migration assay show that in vitro MSC migration was 25 times higher when stimulated with the PBMCs (P = 0.014, Figure 2(d)) even though the cells did not have direct cell-to-cell contact. The directed migration was not found in the negative controls.


Peripheral blood derived mononuclear cells enhance the migration and chondrogenic differentiation of multipotent mesenchymal stromal cells.

Hopper N, Wardale J, Howard D, Brooks R, Rushton N, Henson F - Stem Cells Int (2015)

Cell migration experiment of MSCs measured over 24 hours comparing the effect of PBMC stimulus (n = 4). (a) Wound closure rate measured at 3-hour time point (n = 4) in the scratch assay with direct cell-to-cell contact. (b) The total distance of the wound measured at 12-, 15-, and 24-hour time points is not to scale (n = 4, P < 0.0001) in the scratch assay. (c) The initial cell migration rate at 3-hour time point quantified real time with the xCELLigence system and (d) the total amount of cells migrated at the 24-hour time point quantified with cell index value by xCELLigence.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4309296&req=5

fig2: Cell migration experiment of MSCs measured over 24 hours comparing the effect of PBMC stimulus (n = 4). (a) Wound closure rate measured at 3-hour time point (n = 4) in the scratch assay with direct cell-to-cell contact. (b) The total distance of the wound measured at 12-, 15-, and 24-hour time points is not to scale (n = 4, P < 0.0001) in the scratch assay. (c) The initial cell migration rate at 3-hour time point quantified real time with the xCELLigence system and (d) the total amount of cells migrated at the 24-hour time point quantified with cell index value by xCELLigence.
Mentions: Scratch assay showed that direct cell-to-cell PBMC stimulation increased the wound closure rate by 55.2% at 3 hours, P = 0.002 (Figure 2(a)). The distance of the gap was measured at 12-, 15-, and 24-hour time points (Figure 2(b)) and every time point the PBMC treated wound had smaller scratch area (P < 0.0001). To confirm the findings from the scratch assay and to assess if direct cell-to-cell contact is needed for the chemotactic effect, a second assay based on the Boyden chamber model (xCELLigence) was used to quantify cell migration. The real-time cell migration assay demonstrated that during the first 3 hours the MSC migration was 9 times faster in the PBMC stimulated test group (P = 0.008, Figure 2(c)). The results of the 24-hour migration assay show that in vitro MSC migration was 25 times higher when stimulated with the PBMCs (P = 0.014, Figure 2(d)) even though the cells did not have direct cell-to-cell contact. The directed migration was not found in the negative controls.

Bottom Line: In scratch assay the wound closure was 55% higher after 3 hours in the PBMC stimulated test group (P = 0.002), migration rate was 9 times faster (P = 0.008), and total MSC migration was 25 times higher after 24 hours (P = 0.014).Analysis of MSCs by PCR array demonstrated that PBMCs induced the upregulation of genes associated with chondrogenic differentiation over 15-fold.In conclusion, PBMCs increase both MSC migration and differentiation suggesting that they are an ideal candidate for inclusion in regenerative medicine therapies aimed at cartilage repair.

View Article: PubMed Central - PubMed

Affiliation: Division of Trauma and Orthopaedic Surgery, University of Cambridge, Addenbrooke's Hospital, Hills Road, Cambridge BC2 0QQ, UK.

ABSTRACT
A major challenge in cartilage repair is the lack of chondrogenic cells migrating from healthy tissue into damaged areas and strategies to promote this should be developed. The aim of this study was to evaluate the effect of peripheral blood derived mononuclear cell (PBMC) stimulation on mesenchymal stromal cells (MSCs) derived from the infrapatellar fat pad of human OA knee. Cell migration was measured using an xCELLigence electronic migration chamber system in combination with scratch assays. Gene expression was quantified with stem cell PCR arrays and validated using quantitative real-time PCR (rtPCR). In both migration assays PBMCs increased MSC migration by comparison to control. In scratch assay the wound closure was 55% higher after 3 hours in the PBMC stimulated test group (P = 0.002), migration rate was 9 times faster (P = 0.008), and total MSC migration was 25 times higher after 24 hours (P = 0.014). Analysis of MSCs by PCR array demonstrated that PBMCs induced the upregulation of genes associated with chondrogenic differentiation over 15-fold. In conclusion, PBMCs increase both MSC migration and differentiation suggesting that they are an ideal candidate for inclusion in regenerative medicine therapies aimed at cartilage repair.

No MeSH data available.


Related in: MedlinePlus