Limits...
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

(a) Flow cytometry result of CD90 and CD105 expression in human primary MSCs (n = 6) both normoxia and hypoxia. Results are expressed as mean % of positive cells in the whole population. (b) Multidifferentiation assay for osteogenic (alizarin red), chondrogenic (alcian blue), and adipogenic (oil red O) staining of MSC cultures in both normoxia and hypoxia at day 21 (scale bar 200 nm, n = 4).
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4309296&req=5

fig1: (a) Flow cytometry result of CD90 and CD105 expression in human primary MSCs (n = 6) both normoxia and hypoxia. Results are expressed as mean % of positive cells in the whole population. (b) Multidifferentiation assay for osteogenic (alizarin red), chondrogenic (alcian blue), and adipogenic (oil red O) staining of MSC cultures in both normoxia and hypoxia at day 21 (scale bar 200 nm, n = 4).

Mentions: Adipose tissue-derived MSCs isolated from the infrapatellar fat pad presented CD90 and CD105 cell surface markers typical of an MSC phenotype lacking the CD34/45 expression (measured at passage 4). Hypoxic culture environment similar to that of native cartilage tissue did not have significant effect on MSC progenitor cell population (95.7 ± 3.3% CD90/CD105 positive) when compared to the normoxic culture (84.1 ± 17.0% CD90/CD105 positive) (P = 0.07, Figure 1(a)) although there was a beneficial trend.


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)

(a) Flow cytometry result of CD90 and CD105 expression in human primary MSCs (n = 6) both normoxia and hypoxia. Results are expressed as mean % of positive cells in the whole population. (b) Multidifferentiation assay for osteogenic (alizarin red), chondrogenic (alcian blue), and adipogenic (oil red O) staining of MSC cultures in both normoxia and hypoxia at day 21 (scale bar 200 nm, n = 4).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig1: (a) Flow cytometry result of CD90 and CD105 expression in human primary MSCs (n = 6) both normoxia and hypoxia. Results are expressed as mean % of positive cells in the whole population. (b) Multidifferentiation assay for osteogenic (alizarin red), chondrogenic (alcian blue), and adipogenic (oil red O) staining of MSC cultures in both normoxia and hypoxia at day 21 (scale bar 200 nm, n = 4).
Mentions: Adipose tissue-derived MSCs isolated from the infrapatellar fat pad presented CD90 and CD105 cell surface markers typical of an MSC phenotype lacking the CD34/45 expression (measured at passage 4). Hypoxic culture environment similar to that of native cartilage tissue did not have significant effect on MSC progenitor cell population (95.7 ± 3.3% CD90/CD105 positive) when compared to the normoxic culture (84.1 ± 17.0% CD90/CD105 positive) (P = 0.07, Figure 1(a)) although there was a beneficial trend.

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