Differentiation in vitro of larval cell types from early embryonic cells of Drosophila melanogaster

Development ◽  
1975 ◽  
Vol 33 (1) ◽  
pp. 159-175
Author(s):  
Glen Shields ◽  
Andreas Dübendorfer ◽  
James H. Sang

A variety of cell types develop when cells of 6½-8½ h Drosophila embryos are cultured in an improved medium. Nerve, muscle, fat-body, chitin-secreting, and macrophage-like cells (possibly haemocytes) appear in the first 24 h and mature over the next week. Tracheal, imaginal disc, a second stage of the macrophage-like, and a number of unidentified fibroblastic and epithelial cells appear in the 2nd and 3rd week, following a resumption of cell multiplication. There is some organization of some of the cell types into higher structures.

Development ◽  
1978 ◽  
Vol 45 (1) ◽  
pp. 161-172
Author(s):  
David P. Cross ◽  
James H. Sang

A new procedure is described for the preparation of in vitro cell cultures from individual early gastrulae of Drosophila melanogaster. In these cultures several identifiable cell types differentiate within 24 h (nerve, muscle, fat-body, haemocyte and chitin-secreting): their initial appearance and continuing development over a period of weeks is described. It is proposed that this technique may be used to analyse abnormalities of cellular development in embryonic lethal mutants. Culture in vitro of cells from lethal embryos is seen to have two broad roles: (1) to test the developmental capacity of individual cell types in a situation where they are relatively free from possible deleterious interactions with other cell types and are liberated from the system of the dying embryo, and (2) through the preparation of mixed cultures from normal and mutant embryos, to determine the influence of the presence of wild-type cells on observed abnormalities of a particular cell type.


Development ◽  
1975 ◽  
Vol 34 (2) ◽  
pp. 387-405
Author(s):  
S. A. Iles ◽  
M. W. McBurney ◽  
S. R. Bramwell ◽  
Z. A. Deussen ◽  
C. F. Graham

Mouse eggs were activated with hyaluronidase in vitro and subsequently transferred to the oviduct. In the female reproductive tract they formed morulae and blastocysts which died soon after implantation. Haploid blastocysts were transferred beneath the kidney capsule and here some formed disorganized egg-cylinder structures in a week. Morulae and blastocysts from haploid and diploid parthenogenones were also transferred beneath the testis capsule. Two to four months later the growths which had formed were sectioned. They contained neural tissue, pigment, keratinized epithelium, glandular epithelium, ciliated epithelium, cartilage, bone, muscle, adipose tissue, and haemopoietic tissue. The range of cell types was similar to that produced by fertilized control blastocysts except that the parthenogenones did not form identifiable yolk-sac carcinoma or embryonal carcinomacells. The growths from haploid and diploid parthenogenones in the testis were stained with Feulgen and their DNA content measured. Growths from diploid embryos contained the normal diploid amount of DNA while growths from haploid embryos contained less than this amount. Cell cultures were prepared from the growths. The cells which were investigated contained no Y chromosome, suggesting that they were derived from the embryonic cells rather than the cells of the male host. These cells contained a near diploid chromosome number, although some of them were originally derived from haploid embryos.


Development ◽  
1975 ◽  
Vol 33 (2) ◽  
pp. 487-498
Author(s):  
Andreas Dübendorfer ◽  
Glen Shields ◽  
James H. Sang

Embryos of Drosophila melanogaster, 6–8 h after oviposition, were dissociated and the cells cultured in vitro. Besides larval cell types, imaginal disc cells, assembled and growing in bloated monolayered vesicles, were obtained. The cells of these vesicles become competent to differentiate adult structures when treated with α-ecdysone or ecdysterone in vitro. Recognizable patterns of the adult fly are not formed though. If metamorphosis of imaginal cell vesicles from in vitro-cultures is induced in vivo by transplantation into host larvae of various ages within the third larval instar, recognizable patterns can differentiate provided the host larva does not metamorphose prior to 2 days after transplantation. The frequency of specific patterns in the implants can be increased by providing 9 days of culture in vivo (adult host flies) before metamorphosis. Passage through the third larval instar is not essential for these cells to produce identifiable patterns since culture in adult flies alone can achieve this. The quality of the differentiated pattern is not correlated with the extent of cell proliferation in the cultured tissues. The problem of pattern specification in vitro and in vivo is discussed.


1960 ◽  
Vol 38 (1) ◽  
pp. 871-878 ◽  
Author(s):  
Samuel Dales

To test the effects of anaerobiosis on the rate of multiplication and carbohydrate metabolism of mammalian cells in vitro, cultures of a 'permanent' line, Earle's L strain cells, and of freshly explanted embryonic mouse cells were propagated in the presence and absence of oxygen. Contrary to the findings of several other investigators, our results show that the multiplication of both cell types was depressed by anaerobiosis. Anaerobiosis for at least 7 days, did not, however, bring about unbalanced growth in L cells, nor did it affect their capability to divide rapidly soon after they were returned to aerobic conditions. From the rates of glucose utilization, lactic acid production, and cell multiplication it was estimated that the rate of division in the two cell types studied was proportional to the energy which could be released from either glycolysis or complete oxidation of glucose.


Development ◽  
1998 ◽  
Vol 125 (11) ◽  
pp. 2149-2158 ◽  
Author(s):  
J.S. Britton ◽  
B.A. Edgar

In newly hatched Drosophila larvae, quiescent cells reenter the cell cycle in response to dietary amino acids. To understand this process, we varied larval nutrition and monitored effects on cell cycle initiation and maintenance in the mitotic neuroblasts and imaginal disc cells, as well as the endoreplicating cells in other larval tissues. After cell cycle activation, mitotic and endoreplicating cells respond differently to the withdrawal of nutrition: mitotic cells continue to proliferate in a nutrition-independent manner, while most endoreplicating cells reenter a quiescent state. We also show that ectopic expression of Drosophila Cyclin E or the E2F transcription factor can drive quiescent endoreplicating cells, but not quiescent imaginal neuroblasts, into S-phase. Conversely, we demonstrate that quiescent imaginal neuroblasts, but not quiescent endoreplicating cells, can be induced to enter the cell cycle when co-cultured with larval fat body in vitro. These results demonstrate a fundamental difference in the control of cell cycle activation and maintenance in these two cell types, and imply the existence of a novel mitogen generated by the larval fat body in response to nutrition.


Reproduction ◽  
2014 ◽  
Vol 147 (5) ◽  
pp. D1-D12 ◽  
Author(s):  
R Michael Roberts ◽  
Kyle M Loh ◽  
Mitsuyoshi Amita ◽  
Andreia S Bernardo ◽  
Katsuyuki Adachi ◽  
...  

It is imperative to unveil the full range of differentiated cell types into which human pluripotent stem cells (hPSCs) can develop. The need is twofold: it will delimit the therapeutic utility of these stem cells and is necessary to place their position accurately in the developmental hierarchy of lineage potential. Accumulated evidence suggested that hPSC could develop in vitro into an extraembryonic lineage (trophoblast (TB)) that is typically inaccessible to pluripotent embryonic cells during embryogenesis. However, whether these differentiated cells are truly authentic TB has been challenged. In this debate, we present a case for and a case against TB differentiation from hPSCs. By analogy to other differentiation systems, our debate is broadly applicable, as it articulates higher and more challenging standards for judging whether a given cell type has been genuinely produced from hPSC differentiation.


Development ◽  
1970 ◽  
Vol 23 (1) ◽  
pp. 53-69
Author(s):  
Glen Shields ◽  
James H. Sang

Although attempts have been made over a number of decades to achieve successful tissue culture of Drosophila material, progress has been held back until recently by a lack of basic information on the chemical and physiological characteristics of the haemolymph of the organism necessary for a reasoned formulation of a culture medium. In 1963, Begg & Cruickshank published details of the mineral composition, osmotic pressure and pH of the haemolymph of third instar larvae, and this has provided the basis for a more satisfactory approach, as reflected in an increasing amount of fruitful work since reported. Much of this work has been done with embryonic material. Horikawa & Fox (1964) claimed continuous multiplication of a small type of early embryonic cell; Lesseps (1965) described re-aggregation of dissociated embryonic cells in culture, with possible development of muscle cells, nerve cells and oenocytes in the aggregates.


2017 ◽  
Vol 28 (11) ◽  
pp. 1435-1443
Author(s):  
Lotti Brose ◽  
Justin Crest ◽  
Li Tao ◽  
William Sullivan

Animal cytokinesis involves both actin-myosin–based contraction and vesicle-mediated membrane addition. In many cell types, including early Drosophila embryos, Nuf/FIP3, a Rab11 effector, mediates recycling endosome (RE)–based vesicle delivery to the cytokinesis furrow. Nuf exhibits a cell cycle–regulated concentration at the centrosome that is accompanied by dramatic changes in its phosphorylation state. Here we demonstrate that maximal phosphorylation of Nuf occurs at prophase, when centrosome-associated Nuf disperses throughout the cytoplasm. Accordingly, ectopic Cdk1 activation results in immediate Nuf dispersal from the centrosome. Screening of candidate kinases reveals a specific, dosage-sensitive interaction between Nuf and Polo with respect to Nuf-mediated furrow formation. Inhibiting Polo activity results in Nuf underphosphorylation and prolonged centrosome association. In vitro, Polo directly binds and is required for Nuf phosphorylation at Ser-225 and Thr-227, matching previous in vivo–mapped phosphorylation sites. These results demonstrate a role for Polo kinase in directly mediating Nuf cell cycle–dependent localization.


1960 ◽  
Vol 38 (8) ◽  
pp. 871-878 ◽  
Author(s):  
Samuel Dales

To test the effects of anaerobiosis on the rate of multiplication and carbohydrate metabolism of mammalian cells in vitro, cultures of a 'permanent' line, Earle's L strain cells, and of freshly explanted embryonic mouse cells were propagated in the presence and absence of oxygen. Contrary to the findings of several other investigators, our results show that the multiplication of both cell types was depressed by anaerobiosis. Anaerobiosis for at least 7 days, did not, however, bring about unbalanced growth in L cells, nor did it affect their capability to divide rapidly soon after they were returned to aerobic conditions. From the rates of glucose utilization, lactic acid production, and cell multiplication it was estimated that the rate of division in the two cell types studied was proportional to the energy which could be released from either glycolysis or complete oxidation of glucose.


1977 ◽  
Vol 14 (4) ◽  
pp. 420-425 ◽  
Author(s):  
G. C. Mohanty ◽  
B. S. Rajya

Canine venereal tumor cells were grown in monolayer and tumor tissue fragments were maintained in vitro for 56 days in medium-199. Attachment and cell replication were evident within 17 hours and monolayer was obtained by the tenth day. Growth at first comprised mainly round cells with abundance of cytoplasm and vesicular nuclei, and a few spindle-shaped cells. Later cells became more elongated and like fibroblasts. Cell degeneration started by the 17th day and most were degenerated by the 24th day. Necrosis and depletion of cell population were prominent in tumor explants during the first 2 weeks. Distinct cell multiplication was evident by the 21st day and tumor fragments were repopulated with cells resembling the original tumor by the 56th day. Two distinct morphological cell types were seen: small cells with vesicular, round to oval nuclei and acidophilic cytoplasm; and the large cells with large hyperchromatic nuclei and acidophilic cytoplasm.


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