Isotransplantation of Organ-Cultured Neonatal Pancreas: Reversal of Alloxan Diabetes in the Rat
Isotransplantation of Organ-Cultured Neonatal Pancreas: Reversal of Alloxan Diabetes in the Rat
复制标题
器官培养的新生儿胰腺的同种移植：逆转大鼠四氧嘧啶糖尿病
作者：
O. Hegre;R. J. Leonard;R. V. Schmitt;A. Lazarow
新生大鼠生后4~16天的胰腺在器官培养中培养2~9天。大约10-20个外植体，每个大小为1毫米3(1毫克)，生长在单个微孔过滤器上，放置在由50%马血清和50%鸡胚胎提取物组成的介质的气液界面上。器官培养后，估计为9-20毫克。用胶原酶分离培养的胰岛组织，并将其移植到四氧嘧啶糖尿病受体的腹膜腔内。在8名受者中，有7名患者的糖尿病在移植后11至53天内逆转。动物摄入12-16毫克。培养的胰岛在11-20天内达到正常血糖，9-10 mg。培养的胰岛组织在45~53天内恢复正常。这些动物已经六个多月没有任何症状了。糖尿病逆转后从腹膜腔取材的活体组织含有血管丰富的胰岛，与大量颗粒化的β细胞相比。用线性扫描法对宿主胰腺进行定量分析(在糖尿病逆转后1~2周和4~5个月取材)显示，胰岛β细胞总量为正常值的3%，胰岛素含量为正常值的6%。很少或根本没有观察到宿主β细胞再生的证据。这些研究表明，同种移植前一段时间的器官培养不会损害大鼠胰岛组织逆转四氧嘧啶糖尿病的能力。
Pancreases from neonatal rats four to 16 days postpartum were grown in organ culture for from two to nine days. Approximately 10-20 explants, each measuring 1 mm.3 (1 mg.), were grown on a single Millipore filter placed at the gas-liquid interface of a medium consisting of 50 per cent horse serum and 50 per cent chick embryo extract. Following organ culture, an estimated 9-20 mg. of cultured islet tissue were dissociated with collagenase and isotransplanted into the peritoneal cavity of alloxan-diabetic recipients. In seven of eight recipients the diabetes was reversed between 11 and 53 days of posttransplantation. Animals receiving 12-16 mg. of cultured islet attained normoglycemia in 11-20 days; animals receiving 9-10 mg. of cultured islet tissue recovered between 45 and 53 days. These animals have remained symptom-free for over six months. Biopsies of grafts taken from the peritoneal cavity following reversal of diabetes contained well-vascularized islets compared primarily of heavily granulated beta cells. Quantitative analysis of host pancreases by the linear scanning method (biopsied at one to two weeks and four to five months following reversal of the diabetes) demonstrated that the total beta-cell mass was 3 per cent and the total insulin content was 6 per cent of the normal values. Little or no evidence of regeneration of host beta cells was observed. These studies show that a period of organ culture prior to isotransplantation does not impair the ability of islet tissue to reverse alloxan diabetes in the rat.