Figure 5

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.