Developmental Biology-in vitro chondrogenesis

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Results & Discussion

Upon fixing and staining of our cells, we observed definite cartilage formation in both the wing and leg controls (Figure 1 B1 & Figure 2 C1, table 1). The face cells never adhered properly to the base of the wells. Therefore, when the growth media was pulled out in order to add the paraformaldehyde, the cells unavoidably came out with the media. The hind and wing cells did not come out with the media. They remained in a circular shape on the bottom of the wells in their adhered positions. We took two kinds of pictures. The first was a set of images taken with the inverted scope prior to pulling out the growth media. By doing this, we were hoping to get pictures of chondrogenesis in the face cells without disturbing them. However, we still were not able to visualize the face controls fully because the cells had been scattered before we could take the pictures. We did see three cartilage nodules in the face control, so we know that chondrogenesis did occur (Figure 1 A1). We just do not know how much.

After treatment of each type of cells with retinoic acid (RA), we observed some interesting, but unfortunate occurrences. The RA seemed to settle out of solution and remain on the bottom of the wells in 8 out of the 12 RA-treated wells. This occurrence made it extremely difficult to view the chondrogenesis that may have taken place. We used RA that had been made up for a previous experiment. For future experiments, the RA should be made up fresh, so as to potentially avoid this problem. We took pictures of the wells that were free enough of RA so that they could be visualized (Figure 1 A2, B2, & C2). Generally, we were not able to conclude that RA inhibited chondrogenesis in hind limb buds and stimulated it in wing buds as literature suggests (Figure 2 B2, C2) (Smith 1998). The face results were again indiscernible in the bright field microscope after staining. Prior to fixing and staining, however, the pictures from the inverted scope showed slightly more chondrogenesis than was seen for the hind limb cells

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Last Modified: 2 May 2000

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		Results & Discussion
		Upon fixing and staining of our cells, we observed definite cartilage formation in both the wing and leg controls (Figure 1 B1 & Figure 2 C1, table 1). The face cells never adhered properly to the base of the wells. Therefore, when the growth media was pulled out in order to add the paraformaldehyde, the cells unavoidably came out with the media. The hind and wing cells did not come out with the media. They remained in a circular shape on the bottom of the wells in their adhered positions. We took two kinds of pictures. The first was a set of images taken with the inverted scope prior to pulling out the growth media. By doing this, we were hoping to get pictures of chondrogenesis in the face cells without disturbing them. However, we still were not able to visualize the face controls fully because the cells had been scattered before we could take the pictures. We did see three cartilage nodules in the face control, so we know that chondrogenesis did occur (Figure 1 A1). We just do not know how much. After treatment of each type of cells with retinoic acid (RA), we observed some interesting, but unfortunate occurrences. The RA seemed to settle out of solution and remain on the bottom of the wells in 8 out of the 12 RA-treated wells. This occurrence made it extremely difficult to view the chondrogenesis that may have taken place. We used RA that had been made up for a previous experiment. For future experiments, the RA should be made up fresh, so as to potentially avoid this problem. We took pictures of the wells that were free enough of RA so that they could be visualized (Figure 1 A2, B2, & C2). Generally, we were not able to conclude that RA inhibited chondrogenesis in hind limb buds and stimulated it in wing buds as literature suggests (Figure 2 B2, C2) (Smith 1998). The face results were again indiscernible in the bright field microscope after staining. Prior to fixing and staining, however, the pictures from the inverted scope showed slightly more chondrogenesis than was seen for the hind limb cells