Lim S M, Heng K K, Chong S M, Seah M L, Lim N K, Li S Q
Department of Pathology, National University of Singapore.
Ann Acad Med Singap. 1991 Jul;20(4):472-7.
Porcine fetal pancreases (PFP) obtained from 4 pregnant sows were pooled, minced into 1 mm3 fragments and studied in organ culture for up to 30 days to determine tissue viability and insulin production in vitro. After 7-9 days in culture, some of these explants were transplanted into euglycemic, N:NIH-nu(s) nude recipient mice, and studied histologically over 69 days following grafting. Using RPMI 1640 supplemented with 5% fetal calf serum as the culture medium in 90% air/10% CO2, it was found that explants were viable with insulin production detected in vitro, which was maximal at day 7 (197 +/- 18.9 mU/L, n = 11), and gradually declined thereafter. By 22 days, insulin levels were less than 60.1 +/- 28.5 mU/L (n = 6). Histology of the explants showed viable tissue with evidence of mitoses present in insulin-positive cells at day 16 in vitro. Beyond this time, tissue viability diminished. Explants transplanted into euglycemic nude mice did not undergo rejection during the observation period of 69 days. Grafts remained viable with evidence of an increase in mitotic activity in the endocrine tissue on immunoperoxidase staining. These preliminary investigations confirm that pancreatic explants from porcine fetuses can be maintained in culture for up to 16 days. Such explants, when transplanted under the kidney capsule of euglycemic, nude mice, did not undergo necrosis, but remained viable, with evidence of mitoses in the islet tissue.
从4头怀孕母猪获取的猪胎儿胰腺(PFP)被汇集起来,切碎成1立方毫米的碎片,并在器官培养中研究长达30天,以确定体外组织活力和胰岛素分泌。培养7 - 9天后,将其中一些外植体移植到血糖正常的N:NIH-nu(s)裸受体小鼠体内,并在移植后69天进行组织学研究。使用补充有5%胎牛血清的RPMI 1640作为培养基,在90%空气/10%二氧化碳环境中培养,发现外植体具有活力,体外可检测到胰岛素分泌,在第7天达到最大值(197±18.9 mU/L,n = 11),此后逐渐下降。到第22天,胰岛素水平低于60.1±28.5 mU/L(n = 6)。外植体的组织学显示,体外培养第16天时,组织有活力,胰岛素阳性细胞中有有丝分裂的迹象。超过这个时间,组织活力下降。移植到血糖正常的裸鼠体内的外植体在69天的观察期内未发生排斥反应。移植物保持活力,免疫过氧化物酶染色显示内分泌组织中有丝分裂活性增加。这些初步研究证实,猪胎儿胰腺外植体可在培养中维持长达16天。这种外植体移植到血糖正常的裸鼠肾包膜下时,未发生坏死,而是保持活力,胰岛组织中有有丝分裂的迹象。