Hendrix M J, Seftor E A, Seftor R E, Fidler I J
Department of Anatomy, University of Arizona College of Medicine, Tucson.
Cancer Lett. 1987 Dec;38(1-2):137-47. doi: 10.1016/0304-3835(87)90209-6.
This paper presents a more reliable model for studying the extent of tumor cell migration and invasion in vitro. Polycarbonate filters were uniformly coated with a reconstituted basement membrane material and allowed to dry; each filter measured 0.035 mm in thickness when hydrated with media. Subsequently, the membrane-coated filters were suspended in Membrane Invasion Culture System (MICS) chambers, and high (A375M) and low (A375P) metastatic variants of human melanoma cells were seeded onto the membranes and allowed to incubate for 3 days. At the end of this period, cells were examined morphologically, and the invasive cells of both metastatic variants were collected, stained and counted microscopically. The tumor cells could be seen attached to the reconstituted basement membrane, buried within it, and forming colony-like aggregates in the matrix. It was determined that approximately twice as many A375M cells invaded the artificial biological matrix compared with the A375P cells (P less than 0.0005). Substantially more cells from each variant invaded uncoated polycarbonate filters, thus indicating the selective barrier imposed by the Matrigel. The utilization of such a reconstituted matrix for in vitro invasion studies allows one the opportunity to examine tumor cell attachment, migration and invasion of a uniform matrix over an extended period of time.
本文提出了一种更可靠的模型，用于研究肿瘤细胞在体外迁移和侵袭的程度。将聚碳酸酯滤膜均匀地涂上重组基底膜材料并使其干燥；当用培养基水化后，每个滤膜的厚度为0.035毫米。随后，将涂有膜的滤膜悬浮在膜侵袭培养系统（MICS）小室中，将人黑色素瘤细胞的高转移（A375M）和低转移（A375P）变体接种到膜上，并使其孵育3天。在此期间结束时，对细胞进行形态学检查，并收集两种转移变体的侵袭细胞，进行染色并在显微镜下计数。可以看到肿瘤细胞附着在重组基底膜上，埋入其中，并在基质中形成集落样聚集体。结果确定，与A375P细胞相比，侵入人工生物基质的A375M细胞数量大约是其两倍（P小于0.0005）。来自每个变体的更多细胞侵入未涂覆的聚碳酸酯滤膜，从而表明基质胶施加的选择性屏障。利用这种重组基质进行体外侵袭研究，使人们有机会在较长时间内检查肿瘤细胞对均匀基质的附着、迁移和侵袭情况。
