Tait I S, Flint N, Campbell F C, Evans G S
Department of Surgery, Ninewells Hospital and Medical School, Dundee, UK.
Differentiation. 1994 Apr;56(1-2):91-100. doi: 10.1046/j.1432-0436.1994.56120091.x.
A novel method to study the generation of rat small intestinal mucosa, by transplantation of disaggregated postnatal rat small intestinal epithelium is described. Cellular aggregates, comprised of epithelium with attached proliferative cells and closely associated stromal tissue, were isolated from postnatal rat small intestine by enzymatic digestion, then grafted immediately to the subcutaneous plane of adult recipients. On graft retrieval after 14 days, 39% of cellular transplants to nude mice, and 84% of cellular transplants to inbred rats had developed into small intestine-like structures. These structures were comprised of a circumferential layer of epithelium surrounding a central mucin filled lumen. This neomucosal layer exhibited well formed crypts and villi, and contained all epithelial stem cell lineages i.e. absorptive enterocytes, goblet cells, Paneth's cells and entero-endocrine cells. Proliferative activity within this neomucosa was confined to crypt regions as in normal postnatal small intestine. Developmental maturation within the regenerated neomucosa was demonstrated by organotypic morphogenesis, i.e. formation of mature crypts and villi, and progressive cytodifferentiation with increased numbers of goblet cells, entero-endocrine cells and Paneth's cells. Altered patterns of brush border enzyme expression further confirmed a temporal progression of development within neomucosal enterocytes. It is concluded that after "extensive" mucosal disaggregation, postnatal small intestinal epithelial progenitor cells retain the capacity for organotypic regeneration of neomucosa when transplanted to ectopic sites in adult recipients. These small aggregates of epithelium and stroma are capable of generating the topographical signals necessary for the three dimensional regeneration of this tissue. Furthermore, the multipotent generative potential of the stem cells within these cellular aggregates is maintained with production of all progeny.
本文描述了一种通过移植新生大鼠小肠上皮细胞解离物来研究大鼠小肠黏膜生成的新方法。通过酶消化从新生大鼠小肠中分离出由附着有增殖细胞的上皮细胞和紧密相连的基质组织组成的细胞聚集体，然后立即将其移植到成年受体的皮下平面。14天后取出移植体，移植到裸鼠的细胞移植体中有39%，移植到近交系大鼠的细胞移植体中有84%发育成小肠样结构。这些结构由围绕中央充满粘蛋白的管腔的上皮细胞圆周层组成。这个新黏膜层表现出形成良好的隐窝和绒毛，并包含所有上皮干细胞谱系，即吸收性肠上皮细胞、杯状细胞、潘氏细胞和肠内分泌细胞。如同正常新生小肠一样，这个新黏膜内的增殖活性局限于隐窝区域。再生新黏膜内的发育成熟通过器官样形态发生得以证明，即成熟隐窝和绒毛的形成，以及杯状细胞、肠内分泌细胞和潘氏细胞数量增加的渐进性细胞分化。刷状缘酶表达模式的改变进一步证实了新黏膜肠上皮细胞内发育的时间进程。结论是，在“广泛”的黏膜解离后，新生小肠上皮祖细胞移植到成年受体的异位部位时，保留了新黏膜器官样再生的能力。这些上皮和基质的小聚集体能够产生该组织三维再生所需的地形信号。此外，这些细胞聚集体内干细胞的多能生成潜力在产生所有后代时得以维持。
