Critical parameters for the isolation of mesenchymal stem cells from umbilical cord blood
Critical parameters for the isolation of mesenchymal stem cells from umbilical cord blood
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DOI：
10.1634/stemcells.22-4-625
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发表时间：
2004-01-01
期刊：
影响因子：
5.2
通讯作者：
Eichler, H
中科院分区：
文献类型：
--
作者：
Bieback, K;Kern, S;Eichler, H
有证据表明，间充质干细胞（MSC）是支持细胞治疗新临床概念的有前途的细胞群。然而，之前从足月分娩的脐带血 (UCB) 中分离 MSC 的尝试要么失败，要么产量低。我们研究了是否可以从健康新生儿的 UCB 中培养具有 MSC 特征和多向分化潜力的细胞，以及是否可以通过最佳培养条件或定义 UCB 质量标准来最大化产量。使用优化的分离和培养条件，在 59 个低容量 UCB 单位中，高达 63% 的细胞被分离出显示出特征性间充质形态和免疫表型（NISC 样细胞）的细胞。这些与来自成人骨髓 (BM) 的对照 MSC 相似。 MSC 样细胞的频率范围为每 1 x 10(8) 单核细胞 (MNC) 0 至 2.3 个克隆。细胞克隆广泛增殖，在八代内至少发生了 20 次群体倍增。此外，成骨和软骨分化表现出与骨髓间充质干细胞相当的多谱系能力。然而，与MSC相比，MSC样细胞对成脂分化的敏感性较低。从UCB中分离MSC样细胞的关键点是从采集到分离的时间小于15小时，净体积大于33 ml，MNC计数大于1 x 10(8) MNC。由于可以从足月UCB捐献中高效分离出MSC样细胞，因此我们将UCB视为额外的干细胞用于实验和潜在临床目的的细胞来源。
Evidence has emerged that mesenchymal stem cells (MSCs) represent a promising population for supporting new clinical concepts in cellular therapy. However, attempts to isolate MSCs from umbilical cord blood (UCB) of full-term deliveries have previously either failed or been characterized by a low yield. We investigated whether cells with MSC characteristics and multilineage differentiation potential can be cultivated from UCB of healthy newborns and whether yields might be maximized by optimal culture conditions or by defining UCB quality criteria.Using optimized isolation and culture conditions, in up to 63% of 59 low-volume UCB units, cells showing a characteristic mesenchymal morphology and immune phenotype (NISC-like cells) were isolated. These were similar to control MSCs from adult bone marrow (BM). The frequency of MSC-like cells ranged from 0 to 2.3 clones per 1 x 10(8) mononuclear cells (MNCs). The cell clones proliferated extensively with at least 20 population doublings within eight passages. In addition, osteogenic and chondrogenic differentiation demonstrated a multi-lineage capacity comparable with BM MSCs. However, in contrast to MSCs, MSC-like cells showed a reduced sensitivity to undergo adipogenic differentiation.Crucial points to isolate MSC-like cells from UCB were a time from collection to isolation of less than 15 hours, a net volume of more than 33 ml, and an MNC count of more than 1 x 10(8) MNCs.Because MSC-like cells can be isolated at high efficacy from full-term UCB donations, we regard UCB as an additional stem cell source for experimental and potentially clinical purposes.