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Increased Haematopoietic Supportive Function of USSC from Umbilical Cord Blood Compared to CB MSC and Possible Role of DLK-1.

Kluth SM, Radke TF, Kögler G - Stem Cells Int (2013)

Bottom Line: Multipotent stromal cells can be isolated from a variety of different tissues in the body.In this study, experiments assessing the haematopoiesis-supporting capacity and molecular biological analyses were conducted and clearly confirmed different properties.Compared to CB MSC, USSC lead to a higher expansion of haematopoietic cells and in addition express significantly higher levels of insulin-like growth factor binding protein 1 (IGFBP1), but lower levels of IGF2.

View Article: PubMed Central - PubMed

Affiliation: Institute for Transplantation Diagnostics and Cell Therapeutics, Heinrich Heine University Medical Center, 40225 Duesseldorf, Germany.

ABSTRACT
Multipotent stromal cells can be isolated from a variety of different tissues in the body. In contrast to stromal cells from the adult bone marrow (BM) or adipose tissue, cord blood (CB) multipotent stromal cells (MSC) are biologically younger. Since first being described by our group, delta like 1 homologue (DLK-1) was determined as a discriminating factor between the distinct cord blood-derived subpopulations: the unrestricted somatic stromal cells (USSC), which lack adipogenic differentiation capacity, and the BM MSC-like CB MSC. In this study, experiments assessing the haematopoiesis-supporting capacity and molecular biological analyses were conducted and clearly confirmed different properties. Compared to CB MSC, USSC lead to a higher expansion of haematopoietic cells and in addition express significantly higher levels of insulin-like growth factor binding protein 1 (IGFBP1), but lower levels of IGF2. The data presented here also indicate that DLK-1 might not be the sole factor responsible for the inhibition of adipogenic differentiation potential in USSC but nevertheless indicates a biological diversity among cord blood-derived stromal cells.

No MeSH data available.


Related in: MedlinePlus

Regulations of DLK-1 in human and mice are not the same. To evaluate mechanisms induced by DLK-1 in human CB MSC upon exposure to DLK-1/Pref1, Western Blot analysis of ERK1/2 and p-ERK1/2 was performed (n = 3 experiments). CB MSC were treated with conditioned media of either the control cells or the DLK-1 overexpressing CB MSC. CB MSC (d0 control) and DLK-1 overexpressing CB MSC (d0 DLK+) were lysed and western blot analysis was performed applying full protein lysates after 30 minutes, 4 hours and 24 hours of incubation with conditioned media, respectively. ERK1/2 was already highly expressed in the non treated cells and remained expressed also in the cells treated. No DLK-1/Pref1 specific upregulation of p-ERK1/2 was detected in the CB MSC. As biological positive control, osteogenic differentiated (day 3 after induction) BM MSC were used. Internal loading control: β-actin.
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Related In: Results  -  Collection


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fig3: Regulations of DLK-1 in human and mice are not the same. To evaluate mechanisms induced by DLK-1 in human CB MSC upon exposure to DLK-1/Pref1, Western Blot analysis of ERK1/2 and p-ERK1/2 was performed (n = 3 experiments). CB MSC were treated with conditioned media of either the control cells or the DLK-1 overexpressing CB MSC. CB MSC (d0 control) and DLK-1 overexpressing CB MSC (d0 DLK+) were lysed and western blot analysis was performed applying full protein lysates after 30 minutes, 4 hours and 24 hours of incubation with conditioned media, respectively. ERK1/2 was already highly expressed in the non treated cells and remained expressed also in the cells treated. No DLK-1/Pref1 specific upregulation of p-ERK1/2 was detected in the CB MSC. As biological positive control, osteogenic differentiated (day 3 after induction) BM MSC were used. Internal loading control: β-actin.

Mentions: To evaluate the phosphorylation status of ERK1/2, CB MSC were treated with cell culture supernatant of CB MSC overexpressing DLK-1 (with extracellular expression of DLK-1 and secreting soluble functional DLK-1 as confirmed by ELISA; Figures 2(b) and 2(c) and ERK1/2 expression was analyzed. For western blot analysis, different time points (30 minutes, 4 hours and 24 hours) were applied to cover the expression kinetics of the protein. Osteogenic induced BM MSC (day 3 after induction) served as positive control for the ERK1/2 activation (Figure 3) since the activation of ERK1/2 is required to induce the expression of matrix producing genes, such as collagens, bone morphogenetic proteins (BMPs) or transforming growth factors (TGFs). Surprisingly, while the non-phosphorylated ERK1/2 was expressed under all conditions in CB MSC, a DLK-1-specific phosphorylation of ERK1/2 was not detected. However, it has to be noted that a reduced adipogenic differentiation was observed in the CB MSC overexpressing the full-length DLK-1 [3].


Increased Haematopoietic Supportive Function of USSC from Umbilical Cord Blood Compared to CB MSC and Possible Role of DLK-1.

Kluth SM, Radke TF, Kögler G - Stem Cells Int (2013)

Regulations of DLK-1 in human and mice are not the same. To evaluate mechanisms induced by DLK-1 in human CB MSC upon exposure to DLK-1/Pref1, Western Blot analysis of ERK1/2 and p-ERK1/2 was performed (n = 3 experiments). CB MSC were treated with conditioned media of either the control cells or the DLK-1 overexpressing CB MSC. CB MSC (d0 control) and DLK-1 overexpressing CB MSC (d0 DLK+) were lysed and western blot analysis was performed applying full protein lysates after 30 minutes, 4 hours and 24 hours of incubation with conditioned media, respectively. ERK1/2 was already highly expressed in the non treated cells and remained expressed also in the cells treated. No DLK-1/Pref1 specific upregulation of p-ERK1/2 was detected in the CB MSC. As biological positive control, osteogenic differentiated (day 3 after induction) BM MSC were used. Internal loading control: β-actin.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3649714&req=5

fig3: Regulations of DLK-1 in human and mice are not the same. To evaluate mechanisms induced by DLK-1 in human CB MSC upon exposure to DLK-1/Pref1, Western Blot analysis of ERK1/2 and p-ERK1/2 was performed (n = 3 experiments). CB MSC were treated with conditioned media of either the control cells or the DLK-1 overexpressing CB MSC. CB MSC (d0 control) and DLK-1 overexpressing CB MSC (d0 DLK+) were lysed and western blot analysis was performed applying full protein lysates after 30 minutes, 4 hours and 24 hours of incubation with conditioned media, respectively. ERK1/2 was already highly expressed in the non treated cells and remained expressed also in the cells treated. No DLK-1/Pref1 specific upregulation of p-ERK1/2 was detected in the CB MSC. As biological positive control, osteogenic differentiated (day 3 after induction) BM MSC were used. Internal loading control: β-actin.
Mentions: To evaluate the phosphorylation status of ERK1/2, CB MSC were treated with cell culture supernatant of CB MSC overexpressing DLK-1 (with extracellular expression of DLK-1 and secreting soluble functional DLK-1 as confirmed by ELISA; Figures 2(b) and 2(c) and ERK1/2 expression was analyzed. For western blot analysis, different time points (30 minutes, 4 hours and 24 hours) were applied to cover the expression kinetics of the protein. Osteogenic induced BM MSC (day 3 after induction) served as positive control for the ERK1/2 activation (Figure 3) since the activation of ERK1/2 is required to induce the expression of matrix producing genes, such as collagens, bone morphogenetic proteins (BMPs) or transforming growth factors (TGFs). Surprisingly, while the non-phosphorylated ERK1/2 was expressed under all conditions in CB MSC, a DLK-1-specific phosphorylation of ERK1/2 was not detected. However, it has to be noted that a reduced adipogenic differentiation was observed in the CB MSC overexpressing the full-length DLK-1 [3].

Bottom Line: Multipotent stromal cells can be isolated from a variety of different tissues in the body.In this study, experiments assessing the haematopoiesis-supporting capacity and molecular biological analyses were conducted and clearly confirmed different properties.Compared to CB MSC, USSC lead to a higher expansion of haematopoietic cells and in addition express significantly higher levels of insulin-like growth factor binding protein 1 (IGFBP1), but lower levels of IGF2.

View Article: PubMed Central - PubMed

Affiliation: Institute for Transplantation Diagnostics and Cell Therapeutics, Heinrich Heine University Medical Center, 40225 Duesseldorf, Germany.

ABSTRACT
Multipotent stromal cells can be isolated from a variety of different tissues in the body. In contrast to stromal cells from the adult bone marrow (BM) or adipose tissue, cord blood (CB) multipotent stromal cells (MSC) are biologically younger. Since first being described by our group, delta like 1 homologue (DLK-1) was determined as a discriminating factor between the distinct cord blood-derived subpopulations: the unrestricted somatic stromal cells (USSC), which lack adipogenic differentiation capacity, and the BM MSC-like CB MSC. In this study, experiments assessing the haematopoiesis-supporting capacity and molecular biological analyses were conducted and clearly confirmed different properties. Compared to CB MSC, USSC lead to a higher expansion of haematopoietic cells and in addition express significantly higher levels of insulin-like growth factor binding protein 1 (IGFBP1), but lower levels of IGF2. The data presented here also indicate that DLK-1 might not be the sole factor responsible for the inhibition of adipogenic differentiation potential in USSC but nevertheless indicates a biological diversity among cord blood-derived stromal cells.

No MeSH data available.


Related in: MedlinePlus