Differentiation of cartilage cells studied by quantitative [3H]thymidine autoradiography in vitro

Erella Livne, Anna Weiss, Michael Silbermann
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引用次数: 5

Abstract

The apical segments of the mandibular condylar cartilage of newborn ICR mice, containing the intact zones of progenitor cells along with a few rows of chondroblasts were initially prelabelled in vitro with [3H]thymidine and were subsequently chased and cultured for as long as eight days. Such explants underwent a process of tissue regeneration, as after three days in culture they reconstituted the original structure of the organ, thus resembling the in vivo appearance of neonatal mandibular condylar cartilage. Cellular proliferation with subsequent differentiation in the regenerating tissue was followed by means of quantitative autoradiography. Immediately after labelling, the autoradiography-positive grains were confined exclusively to progenitor cells. The latter revealed a substantial ability to proliferate in vitro, a fact that was manifested by a progressive increase in the labelling index along the course of the culture. The latter process was followed by cellular differentiation thereby obtaining hypertrophic chondrocytes. The increase in the rate of labelling index and in the total number of [3H]thymidine-labelled cells was significantly correlated with the overall growth of the regenerating explants.

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体外定量[3H]胸腺嘧啶放射自显影研究软骨细胞的分化
新生ICR小鼠下颌髁突软骨的顶端段,包含完整的祖细胞区和几排成软骨细胞,最初在体外用[3H]胸苷预先标记,随后进行长达8天的追赶和培养。这种外植体经历了一个组织再生的过程,在培养三天后,它们重建了器官的原始结构,从而类似于新生儿下颌髁软骨的活体外观。用定量放射自显影法观察再生组织的细胞增殖和随后的分化。标记后,放射自显影阳性颗粒仅局限于祖细胞。后者显示出在体外增殖的实质性能力,这一事实表现为在培养过程中标记指数的逐渐增加。后一过程随后是细胞分化,从而获得肥大的软骨细胞。标记率和[3H]胸腺嘧啶标记细胞总数的增加与再生外植体的整体生长显著相关。
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