ISSN:
0192-253X
Keywords:
F9 ECC
;
Aggregates
;
Embryoid bodies
;
Endoderm
;
Life and Medical Sciences
;
Genetics
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Biology
Notes:
To study the relationship between compaction and differentiation in aggregates of F9 embryonal carcinoma cells, a subline was developed which grows mostly uncompacted in monolayer culture in medium containing a low concentration of calcium (about 0.05 mM). When these cells were trypsinized and cultured in suspension in the same medium, they formed loose, open aggregates, which failed to differentiate into embryoid bodies after exposure to 10 nM retinoic acid, confirming the requirement of compaction for differentiation. If, after culture for 3 days, the uncompacted F9 aggregates were exposed to additional calcium (4 mM), all compacted within an hour. The number of days necessary for aggregates to acquire this ability to compact rapidly was reduced if the monolayer of cells from which the aggregates were derived had been exposed to additional calcium to cause compaction for several days prior to trypsinization and aggregation. Next, treatment of the compacted F9 aggregates with 10 nM retinoic acid was followed by differentiation into embryoid bodies. The number of days required for this was also reduced if the aggregates were formed from previously compacted cells, presumably because compaction of the aggregates occured sooner.The acceleration in compaction and differentiation in aggregates formed from previously compacted cells suggests that some of the proteins important for compaction, which are synthesized in a monolayer of compacted cells, persist through trypsinization and are carried over from monolayer to aggregates. Alternatively, an inhibitor of compaction is decreased in the compacted monolayer. Thus, the process of compaction in its entirety, including its relationship to subsequent differentiation, cannot be studied in aggregates formed from F9 cells grown as usual in the compacted state in monolayer culture. This work provides an alternative system in which aggregation, compaction, and differentiation of F9 cells can be made to occur in stepwise fashion and can be examined separately.
Additional Material:
6 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/dvg.1020100508
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