Liu S C, Parsons C S
J Invest Dermatol. 1983 Jul;81(1):54-61. doi: 10.1111/1523-1747.ep12538884.
Using the combined techniques of Rheinwald and Green, and Liu and Karasek, large quantities of proliferative keratinocytes from involved (PP) or uninvolved (PN) skin of psoriatic patients and from normal skin of non-psoriatic donors (NN) can be obtained. Primary cultures, initiated from a 6-mm punch biopsy, are grown on a 3T3 feeder layer seeded on a collagen surface and fed with Dulbecco's Modified Eagle's Medium containing 20% fetal calf serum, hydrocortisone, epidermal growth factor, and cholera toxin. To test the hypothesis that the mechanism(s) responsible for the abnormal proliferation of psoriatic keratinocytes may be located within the cells themselves, primary cultures are passaged onto collagen surfaces without a feeder layer and maintained with medium plus serum, but no additional supplements, and the growth profiles of the 3 cell populations compared. No difference in morphology among these cells is observed in either primary or passaged cultures. In primary cultures, PP keratinocytes, especially those isolated from active lesions, seem to initiate growth at a slower rate than do PN and NN keratinocytes. The difference in the growth rate, as determined by cell number, DNA content, and mitotic activity, is insignificant among passaged PP, PN, and NN cells. Waves observed in the mitotic index and the metabolic activity of the passaged keratinocytes from all 3 sources suggest that the cells are synchronized during subculturing. The cells have high metabolic and mitotic activities during the first week after subculturing, indicative of an initial population of actively dividing cells. We have not found the characteristic feature of hyperproliferation seen in psoriatic keratinocytes in situ, in the cultured cells; however, it is too early to reach the definite conclusion that the mechanism(s) responsible for psoriasis does not exist in the keratinocyte itself. These 3 cell types may respond differently to agents that either enhance or inhibit cell growth and, by using the culture system outlined in this report, we may study these factors and their potential role in psoriasis.
运用莱茵瓦尔德和格林以及刘和卡拉塞克的联合技术，可以从银屑病患者的患部（PP）或非患部（PN）皮肤以及非银屑病供体的正常皮肤（NN）中获取大量增殖性角质形成细胞。从6毫米打孔活检组织起始的原代培养物，在接种于胶原表面的3T3饲养层上生长，并使用含有20%胎牛血清、氢化可的松、表皮生长因子和霍乱毒素的杜尔贝科改良伊格尔培养基进行培养。为了检验银屑病角质形成细胞异常增殖的机制可能存在于细胞自身内部这一假说，将原代培养物传代至没有饲养层的胶原表面，并使用添加血清的培养基进行维持培养，但不添加其他补充剂，然后比较这3种细胞群体的生长情况。在原代培养或传代培养中，未观察到这些细胞在形态上有差异。在原代培养中，PP角质形成细胞，尤其是从活跃病变部位分离的细胞，似乎比PN和NN角质形成细胞起始生长的速率更慢。通过细胞数量、DNA含量和有丝分裂活性确定的传代PP、PN和NN细胞之间的生长速率差异不显著。在来自所有3种来源的传代角质形成细胞的有丝分裂指数和代谢活性中观察到的波动表明，细胞在传代培养过程中是同步的。在传代培养后的第一周，细胞具有高代谢和有丝分裂活性，表明是一群活跃分裂的初始细胞。我们在培养的细胞中未发现银屑病角质形成细胞原位所见的过度增殖特征；然而，现在就得出导致银屑病的机制不存在于角质形成细胞自身的明确结论还为时过早。这3种细胞类型可能对促进或抑制细胞生长的因子有不同反应，通过使用本报告所述的培养系统，我们可以研究这些因素及其在银屑病中的潜在作用。
