探讨大块组织块贴壁培养法在人脐带间充质干细胞（hUC-MSCs）体外分离培养及规模化制备中的应用价值。

采用该法分离培养hUC-MSCs，通过倒置相差显微镜观察细胞形态与生长状态、CCK-8法检测细胞增殖活性、成骨/成软骨诱导分化实验验证多向分化潜能、流式细胞术检测细胞表面标志物。大块组织块法与传统贴壁法在细胞收获量及各时间点吸光度（A）值上的组间差异采用独立样本t检验。

与传统贴壁法相比，大块组织块法的hUC-MSCs细胞收获量（1.5 × 10⁶比1.0 × 10⁶个/皿）更高，差异有统计学意义（P < 0.05）；两种方法获得的细胞增殖活性相当，倍增时间差异无统计学意义（P > 0.05）。培养5 d见细胞迁移，6 ~ 10 d成纤维样细胞迁出，2周融合率超80 %；P3代细胞增殖活性优，表面标志物符合国际标准，且具备成骨、成软骨分化能力。

该方法操作简便、高效低损，可获得高纯度、高活性hUC-MSCs，具有良好推广价值。

To explore the application value of the adherent culture method of large tissue blocks in the in vitro isolation, culture and large-scale preparation of human umbilical cord mesenchymal stem cells (hUC-MSCs).

The large tissue block method was used to isolate and culture hUC-MSCs. Cell morphology and growth status were observed by an inverted phase-contrast microscope. Cell proliferation was detected using the CCK-8 assay. Multidirectional differentiation potential was assessed through osteogenic/chondrogenic induction experiments. Cell surface markers were identified using flow cytometry.

Compared with the traditional adherent culture method, the harvest of hUC-MSCs was higher by the large tissue block method (1.5 × 10⁶ cells/dish vs 1.0 × 10⁶ cells/dish), with a statistically significant difference (P < 0.05). The proliferative activity of cells obtained by both methods was comparable, and there was no statistically significant difference in doubling time (P > 0.05). Cell migration was observed after 5 days of culture, fibroblast-like cells migrated out from 6 to 10 days, and the fusion rate exceeded 80 % after 2 weeks. The proliferation activity of P3 generation cells was excellent, the surface markers met international standards, and they had the ability to differentiate into osteoblasts and chondrocytes.

The large tissue block method is simple to operate, highly efficient and low-loss, could obtain high-purity and high-activity hUC-MSCs, with good promotion value.