Insler M S, Lopez J G
LSU Eye Center, Louisiana State University Medical Center School of Medicine, New Orleans 70112-2234.
Invest Ophthalmol Vis Sci. 1991 May;32(6):1828-36.
To investigate the ability of extended incubation times to improve the success of endothelial cell transplantation, eight human donor corneas were denuded of their native endothelium, seeded twice during a 1-hr interval with a suspension of cultured infant human corneal endothelial cells, and then incubated for 144 hr under standard conditions. Subsequently the corneas were transplanted into African green monkeys using routine penetrating keratoplasty techniques. Rotational autografts and corneas devoid of endothelial cells served as controls. The seeded corneas appeared hazy at the time of surgery (mean pachymetry 48 hr postoperatively, 0.794 mm). Six corneas (75%) subsequently cleared, yielding a mean corneal thickness of 0.541 +/- 0.040 and 0.554 +/- 0.040 at 6 and 12 postoperative months, respectively. All control eyes showed advanced edema (thickness, greater than 1.0 mm) and developed extensive neovascularization. Clinically, the extended postseeding incubation corneas were observed to clear more rapidly and stabilize their thickness earlier than corneas incubated for only 24-48 hr. Scanning electron microscopy of extended postseeding incubation corneas revealed an intact monolayer of contact-inhibited cells with the hexagonal mosaic typical of corneal endothelium in vivo and improved intercellular contact compared with corneas incubated for only 24-48 hr.
为了研究延长孵育时间能否提高内皮细胞移植的成功率，对8只人供体角膜去除其天然内皮，在1小时内分两次接种培养的婴儿人角膜内皮细胞悬液，然后在标准条件下孵育144小时。随后，使用常规穿透性角膜移植技术将角膜移植到非洲绿猴体内。旋转自体移植角膜和无内皮细胞的角膜作为对照。接种后的角膜在手术时看起来模糊（术后48小时平均测厚，0.794毫米）。6只角膜（75%）随后变清晰，术后6个月和12个月时角膜平均厚度分别为0.541±0.040和0.554±0.040。所有对照眼均出现严重水肿（厚度大于1.0毫米）并发展为广泛的新生血管形成。临床上，观察到接种后延长孵育时间的角膜比仅孵育24 - 48小时的角膜更迅速地变清晰且更早地稳定其厚度。对接种后延长孵育时间的角膜进行扫描电子显微镜检查显示，与仅孵育24 - 48小时的角膜相比，有一层完整的接触抑制细胞单层，具有体内角膜内皮典型的六边形镶嵌结构，且细胞间接触有所改善。
