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Expression of cellular marker molecules and extracellular matrix (ECM) proteins by human umbilical cord artery derived cells (HUCAC). Using indirect immunofluorescence staining, highly positive signals (green) were detected for A ) collagen type I of fresh cultivated cells and B ) collagen type I of cryopreserved cells, E ) collagen type <t>III</t> of fresh cultivated cells and F ) collagen type III of cryopreserved cells. The presence of C ) collagen type I (green) and G ) collagen type III (green) was shown in native human umbilical cord artery walls, serving as a control. Immunohistochemical staining verified the presence of D ) collagen type I (red) and H ) collagen type III (red) in native human umbilical cord artery walls. Using flow cytometry analysis, cellular marker expression of short-term (group A, n = 4) and long-term (group B, n = 4) cryopreserved cells from primary cultures (passage 0) was studied directly after I ) thawing and J ) in passage 3 of recultivation. By comparison, non-cryopreserved fresh cells (n = 3) from I ) primary cultures and J ) passage 3 were analyzed in parallel as a control group Using indirect immunofluorescence staining, highly positive signals (green) were detected for all cellular markers tested such as K ) CD90 (green)/ alpha smooth muscle <t>actin</t> <t>(ASMA)</t> (red) of fresh cultivated cells and L ) CD90 (green)/ ASMA (red) of cryopreserved cells, N ) CD29 of fresh cultivated cells and O ) CD29 of cryopreserved cells, P ) CD105 of fresh cultivated cells and Q ) CD105 of cryopreserved cells. Cell nuclei staining is pictured in blue, present in A-H and K-Q. All studies of marker expression are exemplarily shown for cells of passage 3.
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Expression of cellular marker molecules and extracellular matrix (ECM) proteins by human umbilical cord artery derived cells (HUCAC). Using indirect immunofluorescence staining, highly positive signals (green) were detected for A ) collagen type I of fresh cultivated cells and B ) collagen type I of cryopreserved cells, E ) collagen type III of fresh cultivated cells and F ) collagen type III of cryopreserved cells. The presence of C ) collagen type I (green) and G ) collagen type III (green) was shown in native human umbilical cord artery walls, serving as a control. Immunohistochemical staining verified the presence of D ) collagen type I (red) and H ) collagen type III (red) in native human umbilical cord artery walls. Using flow cytometry analysis, cellular marker expression of short-term (group A, n = 4) and long-term (group B, n = 4) cryopreserved cells from primary cultures (passage 0) was studied directly after I ) thawing and J ) in passage 3 of recultivation. By comparison, non-cryopreserved fresh cells (n = 3) from I ) primary cultures and J ) passage 3 were analyzed in parallel as a control group Using indirect immunofluorescence staining, highly positive signals (green) were detected for all cellular markers tested such as K ) CD90 (green)/ alpha smooth muscle actin (ASMA) (red) of fresh cultivated cells and L ) CD90 (green)/ ASMA (red) of cryopreserved cells, N ) CD29 of fresh cultivated cells and O ) CD29 of cryopreserved cells, P ) CD105 of fresh cultivated cells and Q ) CD105 of cryopreserved cells. Cell nuclei staining is pictured in blue, present in A-H and K-Q. All studies of marker expression are exemplarily shown for cells of passage 3.

Journal: Journal of Translational Medicine

Article Title: Cryopreservation of human vascular umbilical cord cells under good manufacturing practice conditions for future cell banks

doi: 10.1186/1479-5876-10-98

Figure Lengend Snippet: Expression of cellular marker molecules and extracellular matrix (ECM) proteins by human umbilical cord artery derived cells (HUCAC). Using indirect immunofluorescence staining, highly positive signals (green) were detected for A ) collagen type I of fresh cultivated cells and B ) collagen type I of cryopreserved cells, E ) collagen type III of fresh cultivated cells and F ) collagen type III of cryopreserved cells. The presence of C ) collagen type I (green) and G ) collagen type III (green) was shown in native human umbilical cord artery walls, serving as a control. Immunohistochemical staining verified the presence of D ) collagen type I (red) and H ) collagen type III (red) in native human umbilical cord artery walls. Using flow cytometry analysis, cellular marker expression of short-term (group A, n = 4) and long-term (group B, n = 4) cryopreserved cells from primary cultures (passage 0) was studied directly after I ) thawing and J ) in passage 3 of recultivation. By comparison, non-cryopreserved fresh cells (n = 3) from I ) primary cultures and J ) passage 3 were analyzed in parallel as a control group Using indirect immunofluorescence staining, highly positive signals (green) were detected for all cellular markers tested such as K ) CD90 (green)/ alpha smooth muscle actin (ASMA) (red) of fresh cultivated cells and L ) CD90 (green)/ ASMA (red) of cryopreserved cells, N ) CD29 of fresh cultivated cells and O ) CD29 of cryopreserved cells, P ) CD105 of fresh cultivated cells and Q ) CD105 of cryopreserved cells. Cell nuclei staining is pictured in blue, present in A-H and K-Q. All studies of marker expression are exemplarily shown for cells of passage 3.

Article Snippet: Fixed cross sectioned umbilical cord arteries were used to localize HUCAC and served as controls by staining with monoclonal mouse anti-human CD90 (4 μg/ml, Dianova), anti-human fibronectin (5 μg/ml, BD Biosciences) and with polyclonal rabbit anti-human ASMA (25 μg/ml, Abcam), anti-human collagen types I & III (0.5 μg/ml, Acris Antibodies), anti-human elastin (1:50, Calbiochem) for 3 h at 4°C.

Techniques: Expressing, Marker, Derivative Assay, Immunofluorescence, Staining, Control, Immunohistochemical staining, Flow Cytometry, Comparison