Morphological alteration of human embryo cells in vitro by treatment with human leukemic culture fluid.

Content Provider	Semantic Scholar
Author	Osato, Toyoro
Copyright Year	1967
Abstract	Search for hypothetical causative agents of viral nature in human leukemias and lymphomas has recently been carried out extensively by electron microscopy, immunofluorescence, and the application of viral interference phenomenon.' On the basis of our results obtained in the in vitro transformation of mouse cells by Friend murine leukemia virus,2 we have attempted a different approach to the problem by testing the transforming capacity, if any, of the human leukemic materials utilizing in vitro cultured human embryonic cells as a "target." The present paper reports the preliminary findings on the appearance of morphological changes in human embryo cells treated with the cell-free supernates of human leukemic cell cultures. Materials and Methods.-Peripheral blood was collected from eight patients, six males and two females, with acute and chronic myelogenous leukemia. The heparinized blood was allowed to sediment at room temperature for one hour to separate the buffy coat. The cells in the buffy coat were pipetted off and placed in the growth medium to compose a cell suspension with 5.0-10 X 106 viable leukocytes per ml. The culture medium employed was M\lcCoy 5A medium containing 30 per cent fetal calf serum.3 Incubation of the leukemic cells was carried out at 370C in a stationary position.4 The "target" cells which were subjected to the exposure with the cell-free supernates of leukemic cell cultures were the primary cultures of three-month-old human embryos. These cultures were prepared in TD40 flasks using Eagle's niedium supplemented with 10 per cent fetal calf serum. Cell-free supernates of leukemic cell cultures were collected between three and seven days of incubation. Trypsinized primary human embryo cells were suspended in the cell-free supernates from the leukemic cultures and subsequently seeded onto TD40 flasks. After incubation overnight at 370C, culture medium was changed with fresh Eagle's medium containing 10 per cent fetal calf serum. Subsequent medium change was carried out every other day. As controls, trypsinized embryo cells were suspended in McCoy 5A medium -with 30 per cent fetal calf serum or in cell-free supernates from leukemic cell cultures treated at 60'C for 30 minutes. Same number of control cultures were set up simultaneously with each series of experiments. The "heated control" was also included in all cases save for the test with first two leukemic cell cultures, nos. 5 and 6. Results.-Eleven different leukemic cell cultures from five acute and three chronic leukemia patients, five untreated and three treated, were tested for their capability to induce morphological changes of human embryo cells. Proliferative "foci" consisting of epithelioid cells were noted among the fibroblastic monolayers composing the background after approximately four to ten weeks of subculture of primary embryo cells with the leukemic culture fluids. Fine network formation by spindleshaped cells distinct from normal human embryonic fibroblasts was another char-
File Format	PDF HTM / HTML
DOI	10.1073/pnas.57.4.1076
PubMed reference number	5340586
Journal	Medline
Volume Number	57
Issue Number	4
Alternate Webpage(s)	http://www.pnas.org/content/57/4/1076.full.pdf
Alternate Webpage(s)	https://doi.org/10.1073/pnas.57.4.1076
Journal	Proceedings of the National Academy of Sciences of the United States of America
Language	English
Access Restriction	Open
Content Type	Text
Resource Type	Article