Vagnoni K E, Ginther O J, Lunn D P
Department of Medical Science, School of Veterinary Medicine, University of Wisconsin-Madison 53706, USA.
Biol Reprod. 1996 Jan;54(1):219-23. doi: 10.1095/biolreprod54.1.219.
Chorionic girdle cells are a highly invasive subpopulation of trophoblastic cells of the horse conceptus that adhere to the uterine epithelium and begin to invade the endometrium on Days 34-36 (Day 0 = day of ovulation). Just prior to and during invasion (Days 32-36), chorionic girdle cells express high levels of major histocompatibility complex (MHC) I, but expression of this antigen decreases by Days 40-45 and is lost by Day 55. The mechanisms involved in the control of chorionic girdle cell invasion and altered MHCI expression over time are not known. The objective of this study, therefore, was to determine the timing of invasion and the characteristics of MHC expression by girdle cells cultured on Matrigel to determine whether chorionic girdle cell behavior in this environment is similar to the behavior in vivo. Chorionic girdles from four conceptuses were collected on each of Days 30, 31, and 32 and placed in Matrigel invasion chambers for 48 h and in additional duplicate chambers for a time period equivalent to the number of days from the day of collection to Day 36 (6, 5, and 4 days, respectively). After these culture times, the area of the filter covered by invasive cells was determined through use of the software program NIH image. At 48 h, Day 31 and Day 32 girdle cell preparations had invaded Matrigel; but only one Day 30 girdle cell preparation showed invasion into Matrigel, and the extent of invasion was limited. Girdle cells collected on all 3 days had invaded Matrigel by the time equivalent to Day 36. Additionally, chorionic girdles from six conceptuses were collected on Day 34 and placed in Matrigel invasion chambers. Three of these were examined for surface antigen expression of MHCI, MHCII, and a trophoblast-specific antigen (102.1) by immunocytochemistry (ICC) every 48 h to a time point equivalent to Day 44. The remaining three were examined for these same antigens at times equivalent to Days 36, 40, 50, and 60. Invasive Day 34 girdle cells expressed MHCI and trophoblast-specific antigen, but not MHCII, at each time examined (Days 36-60). Neither the timing of invasion nor MHCI expression by chorionic girdle cells on Matrigel mimicked what occurs in vivo. Therefore, it is likely that the in utero environment plays a role in regulating these two characteristics of equine chorionic girdle cells.
绒毛膜带细胞是马胚胎滋养层细胞中具有高度侵袭性的亚群，在排卵日第0天起的第34 - 36天附着于子宫上皮并开始侵入子宫内膜。在侵入之前及侵入期间（第32 - 36天），绒毛膜带细胞高水平表达主要组织相容性复合体（MHC）I，但该抗原的表达在第40 - 45天下降，并在第55天消失。绒毛膜带细胞侵袭及MHC I表达随时间变化的调控机制尚不清楚。因此，本研究的目的是确定在基质胶上培养的绒毛膜带细胞的侵袭时间及MHC表达特征，以确定在这种环境下绒毛膜带细胞的行为是否与体内行为相似。在第30、3l和32天分别从四个胚胎收集绒毛膜带，置于基质胶侵袭小室中48小时，并在另外的重复小室中培养相当于从收集日到第36天的天数（分别为6、5和4天）。在这些培养时间后，通过使用软件程序NIH image确定被侵袭细胞覆盖的滤膜面积。在48小时时，第3l和32天的绒毛膜带细胞制剂已侵入基质胶；但只有一个第30天的绒毛膜带细胞制剂显示侵入基质胶，且侵袭程度有限。在相当于第36天的时间时，所有3天收集的绒毛膜带细胞均已侵入基质胶。此外，在第34天从六个胚胎收集绒毛膜带，置于基质胶侵袭小室中。其中三个每48小时通过免疫细胞化学（ICC）检测MHC I、MHC II和滋养层特异性抗原（102.1）的表面抗原表达，直至相当于第44天的时间点。其余三个在相当于第36、40、50和60天的时间检测这些相同抗原。在每次检测时间（第36 - 60天），侵袭性第34天绒毛膜带细胞表达MHC I和滋养层特异性抗原，但不表达MHC II。绒毛膜带细胞在基质胶上的侵袭时间及MHC I表达均未模拟体内发生的情况。因此，子宫内环境可能在调节马绒毛膜带细胞的这两个特征中发挥作用。
