Harkey M A, Whiteley A H
Department of Zoology, University of Washington, 98195, Seattle, Washington, USA.
The Friday Harbor Laboratories, University of Washington, 98195, Seattle, Washington, USA.
Wilehm Roux Arch Dev Biol. 1980 Jun;189(2):111-122. doi: 10.1007/BF00848500.
Methods are described for isolation and culture of primary mesenchyme cells from echinoid embryos. Ninety-five percentpure primary mesenchyme cells were isolated from early gastrulae ofStrongylocentrotus purpuratus, exploiting the biological segregation of these cells within the blastocoel. When cultured, more than 90% of the isolated cells reached the differentiated state, spicule formation, in synchrony with in vivo controls. Isolated primary mesenchyme cells were cultured with and without various cellular and acellular components of normal embryos in order to study the potential involvement of these components in the morphogenesis of the primary mesenchyme. Our data indicate that: 1. primary mesenchyme cells lack the ability to form the annular pattern of the primary mesenchymal ring autonomously; 2. they autonomously produce spicules of a characteristic morphology that differs from that of embryonic spicules; 3. morphogenesis of the primary mesenchyme is not affected by association with embryonic basal lamina, blastocoel matrix, or loosely aggregated epithelial cells, or by close confinement of each set of primary mesenchyme cells within the blastocoelar space; and 4. reaggregated, tightly associated epithelial cells can promote normal primary mesenchyme ring formation, and modify the primary mesenchyme-intrinsic spicule pattern to produce more normal spicule forms.
本文描述了从海胆胚胎中分离和培养原代间充质细胞的方法。利用这些细胞在囊胚腔中的生物学分离特性，从紫海胆的早期原肠胚中分离出了纯度为95%的原代间充质细胞。培养时，超过90%的分离细胞达到分化状态，即同步形成刺，与体内对照一致。为了研究这些成分在原代间充质形态发生中的潜在作用，将分离的原代间充质细胞与正常胚胎的各种细胞和无细胞成分一起培养或不与这些成分一起培养。我们的数据表明：1. 原代间充质细胞缺乏自主形成原代间充质环环形模式的能力；2. 它们自主产生具有特征形态的刺，这种形态不同于胚胎刺；3. 原代间充质的形态发生不受与胚胎基膜、囊胚腔基质或松散聚集的上皮细胞结合的影响，也不受每组原代间充质细胞在囊胚腔空间内紧密限制的影响；4. 重新聚集、紧密相连的上皮细胞可以促进正常的原代间充质环形成，并改变原代间充质内在的刺模式，以产生更正常的刺形态。
