Autoradiographic Techniques for Measurement of the Labeling Index

Simpson-Herren, Linda

doi:10.1007/978-1-60327-406-7_1

Cited by 8 publications

(3 citation statements)

References 41 publications

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“…This criterion consistently included 10-20% of primary tumor cells. The percentage of labeled cells in each metastatic site was divided by the percentage of labeled cells in the same animars primary tumor to obtain a relative thymidine labeling index (TdR) [27].…”

Section: Measurement Of W256 Tumor Cell Growth Ratementioning

confidence: 99%

Stimulation of bone resorption results in a selective increase in the growth rate of spontaneously metastatic Walker 256 cancer cells in bone

et al. 1992

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To test the hypothesis that bone metastasis is related to the rate of bone remodeling, we have examined the effect of enhanced bone resorption on the growth of spontaneously metastatic Walker 256 (W256) cancer cells. Bone resorption was stimulated in male Fischer rats by injecting Rice H-500 Leydig tumor cells subcutaneously. The resorptive response of the skeleton was confirmed in a pilot study by evaluating parameters of bone morphometry after 4, 7 and 10 days of tumor burden. The distal femoral epiphyses had 35 +/- 10% more osteoclast surface, 83 +/- 11% less osteoblast surface, and 46 +/- 5% less trabecular bone after 10 days of tumor burden, compared to non-tumor-bearing controls. To evaluate the effect of Leydig tumor-induced bone resorption on the growth response of W256 cells, 20 rats were injected intramuscularly with 2 x 10(7) W256 cells, and 20 rats were vehicle-injected. Two days later, 10 rats from each group were injected subcutaneously with Leydig tumor cells. Twelve days after W256/vehicle injection, rats were injected with [3H]thymidine, killed 2 h later, and their femurs, liver, lungs and kidneys were processed for histology. In rats injected with Leydig tumor cells only, enhanced bone resorption was confirmed by a 40 +/- 4% increase in serum calcium concentration, a 48 +/- 8% decrease in trabecular bone content, and a 72 +/- 15% decrease in osteoblast surface, compared with non-tumor-bearing rats. Metastatic W256 cells adjacent to trabecular bone in Leydig tumor-bearing rats had a 56 +/- 18% greater relative [3H]thymidine labeling index (TdR) than did W256 cells in the bones of non-Leydig tumor-bearing rats. The TdRs of W256 cells in the liver, lungs, and kidneys were not affected by Leydig tumor burden. In this model, enhanced bone resorption was associated with the selective growth promotion of metastatic W256 cells in bone, suggesting the existence of a bone-derived factor which is mitogenic to W256 cells.

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Section: Measurement Of W256 Tumor Cell Growth Ratementioning

confidence: 99%

Stimulation of bone resorption results in a selective increase in the growth rate of spontaneously metastatic Walker 256 cancer cells in bone

et al. 1992

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“…A technologist trained in handling radioactive labeled isotopes is required to contain and decontaminate areas exposed to tritiated thymidine properly. Routine emulsion exposure is 1 to 4 weeks or longer (3). Strict quality control must be followed to obtain reliable, interpretable results.…”

Section: Introductionmentioning

confidence: 99%

Detection and Evaluation of Proliferating Cell Nuclear Antigen (PCNA) in Rat Tissue by an Improved Immunohistochemical Procedure

Foley¹

1991

his

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Proliferating cell nuclear antigen (PCNA), an endogenous cell replication marker, was detectedby an improvedimmunohistochemical procedure (super sensitive biotin-streptavidin), in formalin fixed, paraffin embedded tissue. Small intestine, liver, kidney, testis, and esophagus from rats were stained for the detection and evaluation ofPCNA as a suitable marker for cell proliferation. Different patterns of staining that are believed to correlate with the individual phases of the cell cycle (Gl, S, G 2 , M, Go) were recognized. PCNA offers an alternative to established celliabeling techniques for investigating cell proliferation. (The J Histotechnol14:237, 1991)

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“…Thymidine labeling has been used for the study of type I1 cells in primary culture (20,231, but its application in vitro has been limited due to the inability of cultured type I1 cells to complete the cell cycle under traditional culture conditions (5,191. Although well established, thymidine labeling protocols are cumbersome and have limited compatibility with other detection methods (25). Over the past decade, flow cytometric techniques have been developed to circumvent these limitations.…”

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confidence: 99%

Flow cytometric study of the type II pneumocyte cell cycle in vivo and in vitro

Uhal

1994

Cytometry

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The type II pneumocyte cell cycle was studied in vivo and in vitro through bivariate flow cytometric analysis of DNA content vs. incorporated 5–bromo‐2–deoxyuridine (BrdUrd). The cell cycle phase durations, Ts (7. 8 h) and TG2/M (1.1 h), measured in vivo, agreed well with earlier estimates obtained by thymidine labeling. Left unilateral pneumonectomy increased the BrdUrd labeling index of type II cells in the remaining lung from an initial value of 1.9 ± 0. 3% to 4.8 ± 1.0%, but had no effect on Ts or TG2/M in vivo. In both normal and pneumonectomized animals, BrdUrd‐positive cells in vivo rapidly completed mitosis but did not enter a second S‐phase. These results demonstrate that proliferating type II cells do not form a continuously cycling population in the normal or regenerating adult lung. When cell cycle parameters were measured in vitro immediately after type II cell isolation, Ts increased 2‐fold and TG2/M rose up to 10‐fold above the value obtained in vivo. After 2 d of primary culture under customary plating conditions, Ts had returned to the same value as that in vivo, but TG2/M remained elevated. Little variability was observed in the duration of S‐phase within each treatment group. In contrast, type II cells exhibited considerable heterogeneity in the rate of G2/M‐phase traverse, especially in vitro. These data suggest that the inability of adult rat type II pneumocytes to proliferate in primary culture is related to delayed G2/M‐phase transit and imply the existence of pneumocyte subpopulations which differ in susceptibility to growth arrest. © 1994 Wiley‐Liss, Inc.

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Autoradiographic Techniques for Measurement of the Labeling Index

Cited by 8 publications

References 41 publications

Stimulation of bone resorption results in a selective increase in the growth rate of spontaneously metastatic Walker 256 cancer cells in bone

Stimulation of bone resorption results in a selective increase in the growth rate of spontaneously metastatic Walker 256 cancer cells in bone

Detection and Evaluation of Proliferating Cell Nuclear Antigen (PCNA) in Rat Tissue by an Improved Immunohistochemical Procedure

Flow cytometric study of the type II pneumocyte cell cycle in vivo and in vitro

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