Itoh Y, Eguchi G
Dev Biol. 1986 Jun;115(2):353-62. doi: 10.1016/0012-1606(86)90255-1.
Pigmented epithelial cells (PECs) were dissociated from eyes of 8- to 9-day-old chick embryos and were cultured in EdF medium (Eagle's MEM supplemented with dialyzed fetal bovine serum) containing phenylthiourea (PTU) and testicular hyaluronidase (HUase). The PECs rapidly lost melanosomes as they proliferated and dedifferentiated in culture. These dedifferentiated PECs (dePECs) which did not manifest any identifiable specificity could be directed to one of two different differentiated phenotypes; viz., lens or pigment cells, depending upon subsequent culture conditions. Almost all dePECs began to synthesize melanin and redifferentiated to PECs by Day 10 of culture with EdF medium containing ascorbic acid (AsA). In contrast, the sister population of dePECs, when cultured at extremely high cell density with EdF medium containing PTU, HUase and AsA, synthesized delta-crystallin which is specific for lens. This transdifferentiation into lens cells occurred by Day 15 of culture. Using this culture system we are able to produce a homogeneous cell population with the potential for synchronous differentiation into either lens or pigment cell phenotype. The system is useful for studying mechanisms involved in cellular metaplasia.
从8至9日龄鸡胚的眼睛中分离出色素上皮细胞（PEC），并将其培养在含有苯硫脲（PTU）和睾丸透明质酸酶（HUase）的EdF培养基（补充有透析胎牛血清的伊格尔氏MEM培养基）中。PEC在培养中增殖和去分化时迅速失去黑素体。这些未表现出任何可识别特异性的去分化PEC（dePEC）可根据后续培养条件被引导至两种不同的分化表型之一，即晶状体或色素细胞。几乎所有的dePEC在含有抗坏血酸（AsA）的EdF培养基中培养到第10天时开始合成黑色素并重新分化为PEC。相反，当dePEC的姊妹群体在含有PTU、HUase和AsA的EdF培养基中以极高的细胞密度培养时，会合成晶状体特异性的δ-晶状体蛋白。这种向晶状体细胞的转分化在培养第15天时发生。利用这个培养系统，我们能够产生具有同步分化为晶状体或色素细胞表型潜力的同质细胞群体。该系统对于研究细胞化生所涉及的机制很有用。
