Isolation and Characterization of Human Umbilical Cord-derived Mesenchymal Stem Cells from Preterm and Term Infants
Isolation and Characterization of Human Umbilical Cord-derived Mesenchymal Stem Cells from Preterm and Term Infants
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DOI：
10.3791/58806
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发表时间：
2019-01-01
影响因子：
1.2
通讯作者：
Nishimura, Noriyuki
中科院分区：
文献类型：
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作者：
Iwatani, Sota;Yoshida, Makiko;Nishimura, Noriyuki
骨髓间充质干细胞（Mesenchymal stem cells，MSCs）具有巨大的治疗潜力，在生物医学领域引起了越来越多的关注。MSC最初从骨髓（BM）中分离和表征，然后从包括脂肪组织、滑膜、皮肤、牙髓和胎儿附属物如胎盘、脐带血（UCB）和脐带（UC）的组织中获得。MSC是具有以下能力的异质细胞群：（1）在标准培养条件下粘附于塑料，（2）表达CD 73（+）/CD 90（+）/CD 105（+）/CD 45（-）/CD 34（-）/CD 14（-）/CD 19（-）/HLA-DR表型的表面标志物，和（3）向脂肪细胞、骨细胞和软骨细胞的三系分化，如国际细胞治疗学会（ISCT）目前所定义的。尽管骨髓是最广泛使用的MSC来源，但骨髓抽吸的侵入性在伦理上限制了其可及性。骨髓间充质干细胞的增殖和分化能力随着供体年龄的增长而下降。相比之下，从UC获得的胎儿MSC具有诸如旺盛的增殖和分化能力的优点。UC采样没有伦理问题，因为它通常被视为医疗废物。人类UC在妊娠4-8周开始发展，羊膜腔持续生长，并持续生长直至达到50-60 cm长，并且可以在整个新生儿分娩期间分离。为了深入了解难治性疾病的病理生理学，我们使用了来自不同胎龄婴儿的UC源性MSC（UC-MSC）。在本方案中，我们描述了从妊娠19-40周的胎儿/婴儿中分离和表征UC-MSC。
Mesenchymal stem cells (MSCs) have considerable therapeutic potential and attract increasing interest in the biomedical field. MSCs are originally isolated and characterized from bone marrow (BM), then acquired from tissues including adipose tissue, synovium, skin, dental pulp, and fetal appendages such as placenta, umbilical cord blood (UCB), and umbilical cord (UC). MSCs are a heterogeneous cell population with the capacity for (1) adherence to plastic in standard culture conditions, (2) surface marker expression of CD73(+)/CD90(+)/CD105(+)/CD45(-)/CD34(-)/ CD14(-)/CD19(-)/HLA-DR- phenotypes, and (3) trilineage differentiation into adipocytes, osteocytes, and chondrocytes, as currently defined by the International Society for Cellular Therapy (ISCT). Although BM is the most widely used source of MSCs, the invasive nature of BM aspiration ethically limits its accessibility. Proliferation and differentiation capacity of MSCs obtained from BM generally decline with the age of the donor. In contrast, fetal MSCs obtained from UC have advantages such as vigorous proliferation and differentiation capacity. There is no ethical concern for UC sampling, as it is typically regarded as medical waste. Human UC starts to develop with continuing growth of the amniotic cavity at 4-8 weeks of gestation and keeps growing until reaching 50-60 cm in length, and it can be isolated during the whole newborn delivery period. To gain insight into the pathophysiology of intractable diseases, we have used UC-derived MSCs (UC-MSCs) from infants delivered at various gestational ages. In this protocol, we describe the isolation and characterization of UC-MSCs from fetuses/infants at 19-40 weeks of gestation.