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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

Haematopoiesis-supporting capacity of USSC and CB MSC. (a) Representative co-culture of CD34+-cells on feeders of USSC, CB MSC, and CB MSC overexpressing DLK-1 (CB MSCDLK) in multiple independent wells (n = 6). With regards to total cell count, expansion of CD34+-cells and colony-forming units (CFU), co-culture on a USSC-feeder resulted in higher expansion rates than on CB MSC-feeder. Within 14 days, total cell count increased significantly stronger on USSC-feeder (by factor 35.05 ± 3.75) than on CB MSC-feeder (factor 10.09 ± 0.37; P < 0.0001). Expansion of total CD34+-cells also resulted in significantly higher expansion on USSC (3.06 ± 0.08-fold) than on CB MSC (1.57 ± 0.02 fold; P < 0.0001). For colony-forming units, cells cultured on USSC-feeder showed a higher expansion rate on day 10 and 14 (up to 4.39 ± 1.50-fold versus 1.06 ± 0.03-fold; P = 0.1562). Overexpression of DLK-1 in the CB MSC line did not have an influence on expansion rates, as the results for CB MSC and CB MSCDLK were approximately identical. (b) Expression levels of haematopoietic cytokines (IGF-2, IGFBP-1, SCF, SDF-1, THPO, ANGPTL-3) in USSC, CB MSC, and BM MSC were analyzed by real time PCR. The expression of insulin-like growth factor binding protein (IGFBP-1) was highest in USSC (n = 3), while stromal-derived factor 1 (SDF-1) expression was higher in CB MSC (n = 3) and BM MSC (n = 3) compared to USSC. Thrombopoietin (THPO) and angiopoietin-like 3 (ANGPTL-3) were not detectable.
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fig4: Haematopoiesis-supporting capacity of USSC and CB MSC. (a) Representative co-culture of CD34+-cells on feeders of USSC, CB MSC, and CB MSC overexpressing DLK-1 (CB MSCDLK) in multiple independent wells (n = 6). With regards to total cell count, expansion of CD34+-cells and colony-forming units (CFU), co-culture on a USSC-feeder resulted in higher expansion rates than on CB MSC-feeder. Within 14 days, total cell count increased significantly stronger on USSC-feeder (by factor 35.05 ± 3.75) than on CB MSC-feeder (factor 10.09 ± 0.37; P < 0.0001). Expansion of total CD34+-cells also resulted in significantly higher expansion on USSC (3.06 ± 0.08-fold) than on CB MSC (1.57 ± 0.02 fold; P < 0.0001). For colony-forming units, cells cultured on USSC-feeder showed a higher expansion rate on day 10 and 14 (up to 4.39 ± 1.50-fold versus 1.06 ± 0.03-fold; P = 0.1562). Overexpression of DLK-1 in the CB MSC line did not have an influence on expansion rates, as the results for CB MSC and CB MSCDLK were approximately identical. (b) Expression levels of haematopoietic cytokines (IGF-2, IGFBP-1, SCF, SDF-1, THPO, ANGPTL-3) in USSC, CB MSC, and BM MSC were analyzed by real time PCR. The expression of insulin-like growth factor binding protein (IGFBP-1) was highest in USSC (n = 3), while stromal-derived factor 1 (SDF-1) expression was higher in CB MSC (n = 3) and BM MSC (n = 3) compared to USSC. Thrombopoietin (THPO) and angiopoietin-like 3 (ANGPTL-3) were not detectable.

Mentions: However, with regard to the discrimination into USSC and CB MSC, here we demonstrate for the first time that only USSC possess an increased expansion capacity (Figure 4(a)) while CB MSC lead to expansion rates comparable to those achieved on BM MSC-feeder (data not shown). To evaluate the impact of DLK-1, co-culture experiments were also performed with CB MSC overexpressing DLK-1 (CB MSCDLK-1). In detail, application of USSC-feeders resulted in higher maximum fold-expansion rates than CB MSC-feeders with regard to total cell count (35.05 ± 3.75 versus 9.78 ± 0.52, n = 6 each, day 14) as well as CD34+ cell count (3.06 ± 0.08 versus 1.58 ± 0.037, n = 3 each, day 14). Additionally, an efficient expansion of colony-forming units (CFU) was observed only on USSC-feeder (4.39 ± 1.50, n = 2, day 10).


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)

Haematopoiesis-supporting capacity of USSC and CB MSC. (a) Representative co-culture of CD34+-cells on feeders of USSC, CB MSC, and CB MSC overexpressing DLK-1 (CB MSCDLK) in multiple independent wells (n = 6). With regards to total cell count, expansion of CD34+-cells and colony-forming units (CFU), co-culture on a USSC-feeder resulted in higher expansion rates than on CB MSC-feeder. Within 14 days, total cell count increased significantly stronger on USSC-feeder (by factor 35.05 ± 3.75) than on CB MSC-feeder (factor 10.09 ± 0.37; P < 0.0001). Expansion of total CD34+-cells also resulted in significantly higher expansion on USSC (3.06 ± 0.08-fold) than on CB MSC (1.57 ± 0.02 fold; P < 0.0001). For colony-forming units, cells cultured on USSC-feeder showed a higher expansion rate on day 10 and 14 (up to 4.39 ± 1.50-fold versus 1.06 ± 0.03-fold; P = 0.1562). Overexpression of DLK-1 in the CB MSC line did not have an influence on expansion rates, as the results for CB MSC and CB MSCDLK were approximately identical. (b) Expression levels of haematopoietic cytokines (IGF-2, IGFBP-1, SCF, SDF-1, THPO, ANGPTL-3) in USSC, CB MSC, and BM MSC were analyzed by real time PCR. The expression of insulin-like growth factor binding protein (IGFBP-1) was highest in USSC (n = 3), while stromal-derived factor 1 (SDF-1) expression was higher in CB MSC (n = 3) and BM MSC (n = 3) compared to USSC. Thrombopoietin (THPO) and angiopoietin-like 3 (ANGPTL-3) were not detectable.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig4: Haematopoiesis-supporting capacity of USSC and CB MSC. (a) Representative co-culture of CD34+-cells on feeders of USSC, CB MSC, and CB MSC overexpressing DLK-1 (CB MSCDLK) in multiple independent wells (n = 6). With regards to total cell count, expansion of CD34+-cells and colony-forming units (CFU), co-culture on a USSC-feeder resulted in higher expansion rates than on CB MSC-feeder. Within 14 days, total cell count increased significantly stronger on USSC-feeder (by factor 35.05 ± 3.75) than on CB MSC-feeder (factor 10.09 ± 0.37; P < 0.0001). Expansion of total CD34+-cells also resulted in significantly higher expansion on USSC (3.06 ± 0.08-fold) than on CB MSC (1.57 ± 0.02 fold; P < 0.0001). For colony-forming units, cells cultured on USSC-feeder showed a higher expansion rate on day 10 and 14 (up to 4.39 ± 1.50-fold versus 1.06 ± 0.03-fold; P = 0.1562). Overexpression of DLK-1 in the CB MSC line did not have an influence on expansion rates, as the results for CB MSC and CB MSCDLK were approximately identical. (b) Expression levels of haematopoietic cytokines (IGF-2, IGFBP-1, SCF, SDF-1, THPO, ANGPTL-3) in USSC, CB MSC, and BM MSC were analyzed by real time PCR. The expression of insulin-like growth factor binding protein (IGFBP-1) was highest in USSC (n = 3), while stromal-derived factor 1 (SDF-1) expression was higher in CB MSC (n = 3) and BM MSC (n = 3) compared to USSC. Thrombopoietin (THPO) and angiopoietin-like 3 (ANGPTL-3) were not detectable.
Mentions: However, with regard to the discrimination into USSC and CB MSC, here we demonstrate for the first time that only USSC possess an increased expansion capacity (Figure 4(a)) while CB MSC lead to expansion rates comparable to those achieved on BM MSC-feeder (data not shown). To evaluate the impact of DLK-1, co-culture experiments were also performed with CB MSC overexpressing DLK-1 (CB MSCDLK-1). In detail, application of USSC-feeders resulted in higher maximum fold-expansion rates than CB MSC-feeders with regard to total cell count (35.05 ± 3.75 versus 9.78 ± 0.52, n = 6 each, day 14) as well as CD34+ cell count (3.06 ± 0.08 versus 1.58 ± 0.037, n = 3 each, day 14). Additionally, an efficient expansion of colony-forming units (CFU) was observed only on USSC-feeder (4.39 ± 1.50, n = 2, day 10).

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