Aoki Takeshi, Jin Zhenghao, Nishino Nobukazu, Kato Hirohisa, Shimizu Yoshinori, Niiya Takashi, Murai Noriyuki, Enami Yuta, Mitamura Keitaro, Koizumi Tomotake, Yasuda Daisuke, Izumida Yoshihiko, Avital Izhak, Umehara Yutaka, Demetriou Achilles A, Rozga Jacek, Kusano Mitsuo
Division of General and Gastroenterological Surgery, Department of Surgery, School of Medicine, Showa University, Tokyo, Japan.
Transplantation. 2005 Apr 15;79(7):783-90. doi: 10.1097/01.tp.0000156319.47645.3b.
Encapsulated cell therapy might be a promising approach to enable cell transplantation without immunosuppression. This study investigates the viability and hepatic function of hepatocytes encapsulated with alginate/poly-L-lysine in vitro and the effect of the intrasplenic transplantation of cultured encapsulated hepatocytes on survival in 90% hepatectomized rats as a preliminary step toward allogeneic hepatocyte transplantation without immunosuppression.
Rat hepatocytes were isolated and encapsulated using alginate/poly-L-lysine. Encapsulated hepatocytes were cultured for 28 days to measure cell viability, liver function, and morphology. Rats were treated with a 90% partial hepatectomy and then immediately underwent the intrasplenic transplantation of the cultured encapsulated hepatocytes, the capsule alone, or the allogeneic hepatocytes without the capsule. The survival rate, liver function, and cell morphology were assessed after transplantation.
The cultured encapsulated hepatocytes maintained their viability and showed better metabolic activity than day 0 cultured encapsulated hepatocytes. The encapsulated cells strongly expressed albumin and were positive for periodic acid-Schiff staining. Electron microscopy demonstrated that the microencapsulated hepatocytes retained the structural elements of hepatic cytoplasm and nuclei. Intrasplenic transplantation of the encapsulated hepatocytes increased the survival rate and improved the hepatic function. Encapsulated hepatocytes transplanted into rat spleen survived well and retained their hepatic function. Moreover, dramatic liver regeneration was observed 48 hr after transplantation in the group that received intrasplenic transplantations of encapsulated hepatocytes.
The intrasplenic transplantation of cultured encapsulated hepatocytes improved the survival rate of an acute liver failure rat model induced by a 90% partial hepatectomy.
封装细胞疗法可能是一种有前景的方法，可实现无需免疫抑制的细胞移植。本研究在体外研究了用海藻酸盐/聚-L-赖氨酸封装的肝细胞的活力和肝功能，以及培养的封装肝细胞脾内移植对90%肝切除大鼠存活的影响，作为迈向无需免疫抑制的同种异体肝细胞移植的初步步骤。
分离大鼠肝细胞并用海藻酸盐/聚-L-赖氨酸进行封装。将封装的肝细胞培养28天，以测量细胞活力、肝功能和形态。对大鼠进行90%部分肝切除术，然后立即进行培养的封装肝细胞、单独的胶囊或无胶囊的同种异体肝细胞的脾内移植。移植后评估存活率、肝功能和细胞形态。
培养的封装肝细胞保持其活力，并且比第0天培养的封装肝细胞表现出更好的代谢活性。封装细胞强烈表达白蛋白，过碘酸-希夫染色呈阳性。电子显微镜显示微封装的肝细胞保留了肝细胞质和细胞核的结构成分。封装肝细胞的脾内移植提高了存活率并改善了肝功能。移植到大鼠脾脏中的封装肝细胞存活良好并保留了其肝功能。此外，在接受封装肝细胞脾内移植的组中，移植后48小时观察到显著的肝再生。
培养的封装肝细胞的脾内移植提高了由90%部分肝切除术诱导的急性肝衰竭大鼠模型的存活率。
