Abstract
We describe a method for preparing highly enriched cultures of Drosophila myoblasts from a heterogeneous cell population derived from gastrulating embryos. Enriched cultures are prepared by plating this heterogeneous population of cells in medium from which much of the free calcium is chelated by ethylene glycol-bis(beta-aminoethyl ether)N,N,N',N'-tetraacetate (EGTA). Adhesion of myoblasts to tissue culture plastic is better than that of other cell types when plated in this medium. Data concerning cell identity, timing of S phase, and fusion kinetics document the degree of enrichment for myogenic cells and illustrate their synchronous differentiation in vitro.
Full Text
The Full Text of this article is available as a PDF (2.4 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Gerson I., Seecof R. L., Teplitz R. L. Ultrastructural differentiation during embryonic Drosophila myogenesis in vitro. In Vitro. 1976 Sep;12(9):615–622. doi: 10.1007/BF02797459. [DOI] [PubMed] [Google Scholar]
- Lengyel J., Spradling A., Penman S. Methods with insect cells in suspension culture. II. Drosophila melanogaster. Methods Cell Biol. 1975;10:195–208. doi: 10.1016/s0091-679x(08)60738-4. [DOI] [PubMed] [Google Scholar]
- Lindsley D. L., Sandler L., Baker B. S., Carpenter A. T., Denell R. E., Hall J. C., Jacobs P. A., Miklos G. L., Davis B. K., Gethmann R. C. Segmental aneuploidy and the genetic gross structure of the Drosophila genome. Genetics. 1972 May;71(1):157–184. doi: 10.1093/genetics/71.1.157. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pardue M. L., Gall J. G. Nucleic acid hybridization to the DNA of cytological preparations. Methods Cell Biol. 1975;10:1–16. doi: 10.1016/s0091-679x(08)60727-x. [DOI] [PubMed] [Google Scholar]
- Paterson B., Strohman R. C. Myosin synthesis in cultures of differentiating chicken embryo skeletal muscle. Dev Biol. 1972 Oct;29(2):113–138. doi: 10.1016/0012-1606(72)90050-4. [DOI] [PubMed] [Google Scholar]
- Petersen N. S., Riggs A. D., Seecof R. L. A method for establishing cell lines from Drosophila melanogaster embryos. In Vitro. 1977 Jan;13(1):36–40. doi: 10.1007/BF02615502. [DOI] [PubMed] [Google Scholar]
- Seecof R. L., Alléaume N., Teplitz R. L., Gerson I. Differentiation of neurons and myocytes in cell cultures made from Drosophila gastrulae. Exp Cell Res. 1971 Nov;69(1):161–173. doi: 10.1016/0014-4827(71)90321-1. [DOI] [PubMed] [Google Scholar]
- Seecof R. L., Dewhurst S. A. A 5-bromodeoxyuridine-sensitive interval during drosophila myogenesis. Differentiation. 1976 Mar 16;6(1):27–32. doi: 10.1111/j.1432-0436.1976.tb01464.x. [DOI] [PubMed] [Google Scholar]
- Seecof R. L., Dewhurst S. Insulin is a Drosophila hormone and acts to enhance the differentiation of embryonic Drosophila cells. Cell Differ. 1974 Jun;3(1):63–70. doi: 10.1016/0045-6039(74)90041-4. [DOI] [PubMed] [Google Scholar]
- Seecof R. L., Donady J. J., Teplitz R. L. Differentiation of Drosophila neuroblasts to form ganglion-like clusters of neurons in vitro. Cell Differ. 1973 Jul;2(3):143–149. doi: 10.1016/0045-6039(73)90014-6. [DOI] [PubMed] [Google Scholar]
- Seecof R. L., Gerson I., Donady J. J., Teplitz R. L. Drosophilia myogenesis in vitro: the genesis of "small" myocytes and myotubes. Dev Biol. 1973 Dec;35(2):250–261. doi: 10.1016/0012-1606(73)90022-5. [DOI] [PubMed] [Google Scholar]
- Shields G., Dübendorfer A., Sang J. H. Differentiation in vitro of larval cell types from early embryonic cells of Drosophila melanogaster. J Embryol Exp Morphol. 1975 Feb;33(1):159–175. [PubMed] [Google Scholar]
- Shields R., Pollock K. The adhesion of BHK and PyBHK cells to the substratum. Cell. 1974 Sep;3(1):31–38. doi: 10.1016/0092-8674(74)90034-8. [DOI] [PubMed] [Google Scholar]
- Turner D. C., Gmür R., Siegrist M., Burckhardt E., Eppenberger H. M. Differentiation in cultures derived from embryonic chicken muscle. I. Muscle-specific enzyme changes before fusion in EGTA-synchronized cultures. Dev Biol. 1976 Feb;48(2):258–283. doi: 10.1016/0012-1606(76)90090-7. [DOI] [PubMed] [Google Scholar]
- Turner D. C., Maier V., Eppenberger H. M. Creatine kinase and aldolase isoenzyme transitions in cultures of chick skeletal muscle cells. Dev Biol. 1974 Mar;37(1):63–89. doi: 10.1016/0012-1606(74)90170-5. [DOI] [PubMed] [Google Scholar]
- Yaffe D. Retention of differentiation potentialities during prolonged cultivation of myogenic cells. Proc Natl Acad Sci U S A. 1968 Oct;61(2):477–483. doi: 10.1073/pnas.61.2.477. [DOI] [PMC free article] [PubMed] [Google Scholar]