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.