Bone marrow assays

Each cytogenetic test system has advantages and disadvantages. For instance, if a very simple determination of a compound s ability to produce chromosomal aberrations in vivo is desired, a bone marrow assay is probably simplest and least expensive. If, however, genetic hazards to future generations are of primary concern, assays of germ cells would be more appropriate. In in vitro assays of peripheral lymphocytes or cell lines, the cells can be made synchronous or fairly synchronous. 

To maximize human relevance, and due to the lack of species limitations for these assays, it is recommended that human cells be used for all in vitro test systems. With the exception of bone marrow assays, the source of cells should be PBL [peripheral blood leukocytes] from donors prescreened for health, immune reactivity, etc. 

Frieauff, W., and Romagna, F. (1994). Technical aspects of automatic micronucleus analysis in rodent bone marrow assays. Cell Biol Toxicol 10, 283-289. 

In a standard mouse micronucleus bone marrow assay, groups of 21 male Swiss albino (CD-I) mice per dose were injected intraperitoneally with 0, 175, 350 or 700 mg A/-(heptan-4-yl)benzo[d][1,3]dioxole-5-carboxamide (No. 1767)/kg bw. At 24, 36 and 48 h following dose administration, seven mice from each group were killed, and their femoral bone marrow was harvested, fixed and stained. No statistically significant differences were observed in the number of polychromatic erythrocytes with micronuclei between the test groups and the negative control (Pucaj, 2004a). 

In a similar standard mouse micronucleus bone marrow assay, male NMRI BR mice (five per group) were administered aqueous A/-gluconyl ethanolamine (No. 1772) at 0 (negative or positive control) or 2000 mg/kg bw via gavage. Femoral bone marrow was isolated at 24 or 48 h post-administration. Treatment and control mice showed no difference in the ratio of polychromatic to normochromatic erythrocytes. 

Zamith. H.P., Vidal, M.N.P.. Speit, G and Paumgartten, F.J.R. (1993) Absence of genotoxic activity of beta-myrcene in the in vivo cytogenetic bone marrow assay. Braz. J. Med. Biol. Res. 26 93-98. 

If any of the genotoxicity studies gives a positive result, an in vivo genotoxicity study is required (bone marrow assay for chromosomal... 

Metaphase analysis or microiuidc-us assay in rodent bone marrow... 

Mouse bone marrow polychromatic-erythrocyte assay (micronucleus test) Micronuclei B Usha Rani et al. 1980... 

Rassi et al., 2002 Porcine bone marrow cells measure osteoclast formation (TRAP staining) and activity (pit assay) Daidzein, at the same concentration as estradiol, inhibits osteoclast formation and activity via caspase-3. 

Tobe et al., 1997 Pit assay with mouse bone marrow cells and dentine slices Daidzein (IQ-Yio-iom) stimulated pit formation while genistein had no effect at this concentration. 

Kado NY, PA Kuzmicky, G Loarca-Pina, MM Mumtaz (1998) Genotoxicity testing of methyl tertiary-butyl ether (MTBE) in the Salmonella microsuspension assay and mouse bone marrow micronucleus test Mut Res 412 131-138. 

In a micronucleus assay using male B6C3Fj mice dosed with 0, 250, 500, or 1,000 mg/kg diisopropyl methylphosphonate, a small but significant increase in micronuclei were observed at mid- and high-dose levels (DOD 1991a). However, the maximum response was found to be within the laboratory historical control limits. The assay was repeated and the increase in micronuclei was not observed, therefore, it is believed that diisopropyl methylphosphonate did not cause micronuclei induction in this experiment. Diisopropyl methylphosphonate was also negative for the induction of micronuclei in the rat bone marrow after administration of up to 800 mg/kg (DOD 1991b). 

Aniline gave positive responses in the mouse bone-marrow micronucleus assay when administered via ingestion or intraperitoneal injection (Ashby et al. 1991 Westmoreland and Gatehouse 1991). However, the positive responses occurred only at a specific time after administration and at what the authors considered high doses (1,000 mg/kg orally and 300 mg/kg intraperitoneally). 

Ashby, J., D.A.Vlachos, and H.Tinwell. 1991. Activity of aniline in the mouse bone marrow micronucleus assay. Mutat. Res. 263 115—117. 

Parodi, S., M.Taningher, P.Russo, M.Pala, M.Tamaro, and C.Monti-Bragadin. 1982. DNA damage in liver, kidney, bone marrow and spleen of rats and mice treated with commercial and purified aniline as determined by alkaline elution assay and sister chromatid exchange induction. Cancer Res. 42 2277—2283. 

The routine transduction of haematopoietic stem cells has, thus far, proven technically difficult. They are found only in low quantities in the bone marrow, and there is a lack of a suitable assay for stem cells. However, recent progress has been made in this regard, and routine transduction of such cells will likely be achievable within the next few years. 

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