Otsuka H, Yoshida K, Kusano S, Okamura Y
Nihon Sanka Fujinka Gakkai Zasshi. 1987 Feb;39(2):271-8.
To evaluate the effects of drugs on uterine leiomyoma and to clarify the histogenesis of uterine leiomyoma, we studied the establishment of primary culture of cells derived from uterine leiomyoma. Myoma tissues were cut into small pieces and suspended in trypsin. The primary cell culture, as a monolayer, was able to be passaged four times. To confirm that the cultured cells were derived from the myoma, the cells were stained with desmin by the enzyme-labelled antibody method. The cultured cells derived from uterine muscle cells were similarly treated with desmin staining. The results confirmed the morphological similarity between the two groups of cells. To confirm that the cultured cells were myoblast, fibroblasts derived from the myoma were cultured selectively. The cells that resembled myoblasts, morphologically appeared spindle-shaped. The cells that resembled fibroblast, appeared polygonal and extended over the bottom of the culture flask. The growth curve of cultured myoma cells (8.0 X 10(4) cells in 0.2 ml) in Petri dishes (60 X 15 mm) revealed logarithmic proliferation after about 5 days. The colony formation of cultured myoma cells (2 X 10(5) cells in 0.2 ml) in culture flasks (25 cm2 in the area of the base) morphologically appeared to have an irregular border and had many independent scattered cells around the colony when the medium was renewed twice a week. The myoma cells (8.8 X 10(4) in 0.2 ml) in the logarithmic phase were cultured, fixed after 12 days when the medium was not renewed, and stained with crystal-violet. Morphologically, the colony had a comparatively regular border, was round, and each cell in the colony was epithelioid. The plating efficiency was 0.07%.(ABSTRACT TRUNCATED AT 250 WORDS)
为评估药物对子宫肌瘤的作用并阐明子宫肌瘤的组织发生,我们研究了子宫肌瘤来源细胞原代培养的建立。将肌瘤组织切成小块并悬浮于胰蛋白酶中。原代细胞培养作为单层培养,能够传代4次。为证实培养的细胞来源于肌瘤,采用酶标抗体法用结蛋白对细胞进行染色。对子宫肌细胞来源的培养细胞同样进行结蛋白染色处理。结果证实两组细胞在形态上相似。为证实培养的细胞是成肌细胞,对肌瘤来源的成纤维细胞进行选择性培养。形态上类似成肌细胞的细胞呈梭形。类似成纤维细胞的细胞呈多边形并伸展于培养瓶底部。培养的肌瘤细胞(0.2 ml中含8.0×10⁴个细胞)在培养皿(60×15 mm)中的生长曲线显示,约5天后呈对数增殖。培养的肌瘤细胞(0.2 ml中含2×10⁵个细胞)在培养瓶(底部面积25 cm²)中的集落形成,当每周更换两次培养基时,形态上边界不规则,集落周围有许多独立分散的细胞。对数期的肌瘤细胞(0.2 ml中含8.8×10⁴个)进行培养,在不更换培养基的情况下培养12天后固定,并用结晶紫染色。形态上,集落边界相对规则,呈圆形,集落中的每个细胞呈上皮样。接种效率为0.07%。(摘要截断于250字)