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. 2020 Jan 13;10:156. doi: 10.1038/s41598-019-57057-8

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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(a) Schematic representation of layer-by-layer technique utilized for encapsulating single cells using the polymer chitosan and alginate. (b) Confocal Laser Scanning Microscopy (CLSM): images of the CA-HSCs (i) DIC, (ii) DAPI, (iii) chitosan-FITC layer, (iv) alginate-RBITC layer, (v) merge and (vi) DIC merge (objective: 60X, Scale: 10 µm); DAPI: 4′, 6-diamidino-2-phenylindole, UV Filter, Excitation/Emision: 358/461 nm; FITC: Fluorescein isothiocyanate, Blue filter, Excitation/Emission: 490/525 nm; RBITC: Rhodamine-β-isothiocyanate, Green Filter, Excitation/Emission: 543/569 nm. (c) Scanning electron microscopy (SEM): images of HSCs and CA-HSCs (Scale: 2 µm). (d) Bar graph representing the results of the calcein-AM/ethidium homodimer-1 live/dead assay (n = 3, results as mean ± SD, *P ≤ 0.05, ^#P ≤ 0.01). (e) Template stability studies: line graph obtained after incubation of templated cells in 100% serum at different time intervals. CA-HSCs were incubated in 100% serum for 2, 4, 6 and 8 hrs to access the degradation. Cells started to lose their template after six hour ((n = 3, results as mean ± SD,*P ≤ 0.05, ^#P ≤ 0.01).