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仔猪脂肪来源干细胞的体外神经元样分化

Neuron-like differentiation of adipose-derived stem cells from infant piglets in vitro.

作者信息

Huang Tingting, He Dansha, Kleiner Gary, Kuluz John

机构信息

University of Miami School of Medicine, Miami, Florida, USA.

出版信息

J Spinal Cord Med. 2007;30 Suppl 1(Suppl 1):S35-40. doi: 10.1080/10790268.2007.11753967.

Abstract

BACKGROUND/OBJECTIVE: Adipose-derived stem cells (ADSCs) are mesenchymal stem cells (MSCs) that can be extracted from adipose tissue and obtained by a less invasive method and in larger quantities compared with bone marrow-derived MSCs. The objective of this study was to harvest ADSCs from piglets and to explore their neuronal differentiation potential.

METHODS

Adipose tissue from piglet facial or abdominal fat was digested with collagenase type XI, followed by filter and centrifugation; the isolated adipose stromal cells were cultured in dishes. MSC markers were measured by flow cytometry; 2 to 5 passage cells were used for in vitro differentiation. Adipogenic, chondrogenic, osteogenic, and neuronal differentiation was induced by incubation of the ADSCs with different induction media.

RESULTS

ADSCs were easily expanded to beyond 15 passages, maintaining the undifferentiated state and exhibiting MSC characteristics and markers CD29, CD44, and CD90. ADSCs differentiated into other mesodermal cells including adipocytes, chondrocytes, and osteocytes. These cells were induced to differentiate into neuron-like cells as evidenced by neuronal morphology and the presence of neuronal markers including microtubule-associated protein 2, neuronal nuclear antigen, and beta-tubulin III.

CONCLUSIONS

ADSCs can be readily obtained from a small amount fat tissue and expanded in culture. Adipose tissue may be an alternative source of stem cell therapy for nervous system injury.

摘要

背景/目的:脂肪来源干细胞(ADSCs)是一种间充质干细胞(MSCs),可从脂肪组织中提取,与骨髓来源的MSCs相比,获取方法侵入性较小且数量更多。本研究的目的是从小猪中获取ADSCs,并探索其向神经元分化的潜能。

方法

用XI型胶原酶消化小猪面部或腹部脂肪组织,然后过滤并离心;将分离出的脂肪基质细胞接种于培养皿中培养。通过流式细胞术检测MSC标志物;使用2至5代细胞进行体外分化。将ADSCs与不同诱导培养基孵育,诱导其向脂肪、软骨、骨和神经元方向分化。

结果

ADSCs易于传代超过15代,保持未分化状态,并表现出MSC特性及标志物CD29、CD44和CD90。ADSCs可分化为其他中胚层细胞,包括脂肪细胞、软骨细胞和骨细胞。这些细胞被诱导分化为神经元样细胞,表现为神经元形态,并存在包括微管相关蛋白2、神经元核抗原和β-微管蛋白III在内的神经元标志物。

结论

ADSCs可从小量脂肪组织中轻易获取并在培养中扩增。脂肪组织可能是神经系统损伤干细胞治疗的替代来源。

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