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Fig. 4 | Stem Cell Research & Therapy

Fig. 4

From: Cryopreserved clumps of mesenchymal stem cell/extracellular matrix complexes retain osteogenic capacity and induce bone regeneration

Fig. 4

Protective role of type I collagen (COLI) against cryopreservation-induced damage in MSC spheroids and C-MSCs. a,b Clumps of mesenchymal stem cell/extracellular matrix complexes (C-MSCs) were treated with or without collagenase for 15 min before cryopreservation. d,e MSC spheroids were generated with growth medium in the presence or absence of 50 μg/mL l-ascorbic acid. Histological analysis of cell constructs before cryopreservation (a,d) or at 5 days after thawing (b,e). Serial sections were stained with hematoxylin and eosin (HE) and TUNEL as indicated. Scale bars = 200 μm. Magnified images of the boxed regions are shown in the panels to the immediate right. In addition, the samples before cryopreservation were immunostained with an anti-type I collagen antibody (green). Nuclei and F-action were counterstained with 4′,6-diamidino-2-phenylindole (DAPI; blue) and Alexa594-conjugated phalloidin (red), respectively. Scale bars = 200 μm. c,f The percentage of TUNEL-positive apoptotic cells in C-MSCs (c) or MSC spheroids (f) 5 days after thawing. Values represent means ± SD of four cultured cell constructs. Similar results were obtained from three independent experiments. *p < 0.05, **p < 0.01, by t test

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