Abstract
Growth in soft-agar bilayer cultures of human tumour cells derived from 4 in vitro continuous cell lines, from 21 xenografts carried in athymic mice, and from 197 samples of fresh human solid tumours of various histologic types was analyzed by computer-assisted image analysis. Replicate cultures for each specimen were assessed on successive days of incubation for the number and volume of growth units within multiple size categories. Our results confirm the recent finding of others that there is an upper limit of approximately 10(9) microns 3 to the cumulative growth unit volume obtainable in a 2 ml bilayer soft agar culture system. Since this upper limit to the carrying capacity of the closed culture system exists, the extent of growth within the cultures is determined in a fundamental way by the cumulative volume of growth units initially inoculated into cultures. A growth index of greater than or equal to 16-fold was only seen when initial cumulative growth unit volume was less than 10(7) microns 3 per culture dish. Computer-assisted volume analysis (CAVA) appears to be a useful quantitative method to study the growth of human tumour cells in soft agar cultures.
Highlights
During the past 5 years, we have studied samples from over 6000 different human solid tumours of various histologic types using this assay methodology
We have developed a number of methodologic modifications to eliminate or control for some of the technical problems associated with primary soft agar cultures prepared using the. basic Hamburger-Salmon technique (Hamburger & Salmon, 1977; Alley et al, 1982; Alley & Lieber, 1984a b)
While the basal layer of cultures was prepared with 0.5% agar, the cellular layer was formulated with 0.3% Seaplaque agarose
Summary
Digestion, and filtration have been described previously (Alley & Lieber, 1984a). Disaggregated cells were filtered by gravity or minimal pressure through one or more 100pm pore size filtration units. Cell suspensions were filtered through a 70pm pore size filtration unit just prior to microscopic examination. Suspensions exhibiting cellular aggregates >60 pm in diameter were filtered one additional time through a 48pm pore size filtration unit prior to cell count. Final suspensions for cell culture were prepared only by dilution of previously filtered suspensions. The following primary human tumour types were studied: colectoral carcinoma, 59 specimens; lung carcinoma, 36; ovarian carcinoma, 23; renal cell carcinoma, 15; breast carcinoma, 7; melanoma, 4; miscellaneous tumour types, 53. Subsequent xenograft tumour cell preparation was carried out exactly as described above for primary human tumour material
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
