The mouse bone marrow cells were cultured in liquid culture system to investigate the correlation of cytochemical changes and the number of colony forming unit granulocyte/macrophage(CFUGM) and colony-forming unit megakaryocyte(CFU-Meg) colonies with cell maturation and platelet. formation:
The following results are obtained;
1. Total colony counts are markedly increased during 4-6 days of culture and reach the maximum number by´7th day (262.33 192.33/ml). The number of cells are increased in each colonies without increase of´ total colony number after, 5 days.
2. The mean ratio of acetylcholinesterase positive megakaryocyte colonies are 13.70 + 7.04 To of total-colonies formed with no remarkable changes during 9 days culture period. The number, of´ megakaryocyte colonies reach maximum level by 7th day (31.37 25.73/ml), while that of non megakaryocytic colonies reach maximum level by 9th day (230.51 87.90/ml).
3. Platelet count in medium are 5,000 3,490/ml with no remarkable changes during 9 days of´ culture periods.
4.. In acetylcholinesterase positive megakaryocytic colonies, the PAS stain show weak positive; in few megakaryocyte by 3rd day, however about 40% of megakaryocytes show rather strong positivity by 7th day. The positivity of non-specific esterase and acid phosphatase are less than 10´% until´ 9th day of culture. Specific esterase positivity was not found during 9 days period of culture.
5. Non-specific esterase and acid phosphatase stain show lower positiviey (1170, 5 %) than PAS and specific esterase stain (21 To ; 55 To) in non megakaryocyte colonies.
In summary; the number of CFU-GM and CFU-Meg colonies in liquid culture are increased gradually and reached maximum by 7th day, forming large colonies from 5th days on. In megakaryocytic colonies, no evidence of platelet formation were observed during 9 days period of culture. It is suggesed that the majority of CFU-GM colonies in liquid culture are mixed colonies according to the cytochemical stainabilities.
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