Human cell-sources for heart valve tissue engineering.

Human cell-sources for heart valve tissue engineering. - The HVTE Cell-source study

Status

Active, not recruiting

Phases

Unknown

Study type

Observational

Source

NL-OMON

Registry ID

NL-OMON33239

Enrollment

Registered

2009-09-22

Start date

2009-10-08

Completion date

Unknown

Last updated

2024-08-12

For informational purposes only — not medical advice. Sourced from public registries and may not reflect the latest updates. Terms

Conditions

not applicable

Interventions

None listed

Eligibility

Age

18 Years to 99 Years

Inclusion criteria

Inclusion criteria: Patients that must undergo on pump coronary artery bypass-grafting including a venous graft.

Exclusion criteria

Exclusion criteria: None

Design outcomes

Primary

Measure	Time frame
The output of this study is composed of several components: - Determination of expansion rate: which celltype can be expanded in the shortest period of time to reach clinically relevant numbers. - Determination of cell phenotype: are the isolated cells still progenitors after in vitro expansion, or did they differentiate into a more specialized celltype? Known markers for the cells of interest are: o MFs: vimentin and a-smooth muscle actin. o Cardiac auricle: c-kit, Sca1, GATA4 o Bone marrow: CD73, CD90, CD105 o Blood: CD14 For this, we will use PCR, immunocytochemistry and FACS-analysis. - Determination of ability to produce matrix. To be suitable for heart valve tissue engineering, cells need to be able to produce sufficient amounts of collagen. In our investigation, we will analyze this on mRNA and protein levels by investigating expression of marker protein Hsp47 and expression of collagen type 1 and 3. This will be done by PCR, immunocytochemistry and western blotting. - Determine ability to produce proteins involved in matrix remodeling. Although the aortic valve should be able to resists high blood pressure, excessive production of matrix can lead to stiff (fibrotic or stenotic) valves. The cells therefore need to be able to produce proteins that can degrade matrix and proteins that inhibit matrix degradation. This way, excessive matrix can be removed to prevent fibrosis, while it can be maintained at locations that bear the heaviest loads. To evaluate ability to remodel matrix, we will look at expression of proteases (MMP1 and 2) and their tissue inhibitors (TIMP1 and 2) on both mRNA and protein level, using PCR, zymography and western blotting. - Finally, we aim to seed expanded cells on a small strip of a degradable carrier, to test the ability of the cells to make a strong tissue strip. Strength of the tissue will be tested using straining-assays and several histochemical analyses to evaluate production and deposition of colla	—

Measure

Time frame

The output of this study is composed of several components: - Determination of expansion rate: which celltype can be expanded in the shortest period of time to reach clinically relevant numbers. - Determination of cell phenotype: are the isolated cells still progenitors after in vitro expansion, or did they differentiate into a more specialized celltype? Known markers for the cells of interest are: o MFs: vimentin and a-smooth muscle actin. o Cardiac auricle: c-kit, Sca1, GATA4 o Bone marrow: CD73, CD90, CD105 o Blood: CD14 For this, we will use PCR, immunocytochemistry and FACS-analysis. - Determination of ability to produce matrix. To be suitable for heart valve tissue engineering, cells need to be able to produce sufficient amounts of collagen. In our investigation, we will analyze this on mRNA and protein levels by investigating expression of marker protein Hsp47 and expression of collagen type 1 and 3. This will be done by PCR, immunocytochemistry and western blotting. - Determine ability to produce proteins involved in matrix remodeling. Although the aortic valve should be able to resists high blood pressure, excessive production of matrix can lead to stiff (fibrotic or stenotic) valves. The cells therefore need to be able to produce proteins that can degrade matrix and proteins that inhibit matrix degradation. This way, excessive matrix can be removed to prevent fibrosis, while it can be maintained at locations that bear the heaviest loads. To evaluate ability to remodel matrix, we will look at expression of proteases (MMP1 and 2) and their tissue inhibitors (TIMP1 and 2) on both mRNA and protein level, using PCR, zymography and western blotting. - Finally, we aim to seed expanded cells on a small strip of a degradable carrier, to test the ability of the cells to make a strong tissue strip. Strength of the tissue will be tested using straining-assays and several histochemical analyses to evaluate production and deposition of colla

—

Countries

Netherlands

Outcome results