Open Access

ARTICLE

Characterization of three-dimensional multipotent adipose-derived stem cell spheroids

HONGYANG LI^1,2, CHAN WANG⁴, SHIWEI LIU¹, YONGLONG GUO^1,3,4,*, JIANSU CHEN^1,3,4,*
1 Ophthalmology Department, First Affiliated Hospital of Jinan University, Guangzhou, 510632, China
2 Ophthalmology Department, Guangdong Second Provincial General Hospital, Guangzhou, 510310, China
3 Institute of Ophthalmology, Medical College, Jinan University, Guangzhou, 510632, China
4 Key Laboratory for Regenerative Medicine of Ministry of Education, Jinan University, Guangzhou, 510632, China
* Corresponding Authors: YONGLONG GUO. Email: ; JIANSU CHEN. Email:
(This article belongs to this Special Issue: Mesenchymal Stem Cells, Secretome and Biomaterials: Regenerative Medicine Application)

BIOCELL 2022, 46(7), 1705-1716. https://doi.org/10.32604/biocell.2022.018442

Received 25 July 2021; Accepted 29 October 2021; Issue published 17 March 2022

Abstract

Human adipose stem cells (hADSCs) are reliable sources for cell therapy. However, the clinical applications are limited by the decrease in activity during in vitro culture. We used a knockout serum replacement (KSR) medium, Eppendorf (EP) tube culture, and a simulated microgravity (SMG) culture system to establish hADSC spheroids. We found that hADSCs aggregated and formed spheroids in the KSR culture medium. The EP tube culture method revealed many biological cell characteristics, such as good cell viabilities, rough surfaces, polar growth, fusion phenomenon, and injectability. The findings show its advantages for hADSCs spherical cultures. When cultured in SMG, hADSC spheroids produced large-scale spheroids. Additionally, confocal examination and viability assays revealed that SMG-cultured hADSC spheroids had higher cell viabilities and looser spherical structures, relative to those cultured in EP tubes. hADSC spheroids in static EP tube culture had tighter structures and more dead cells with rough and irregular surfaces, while hADSC spheroids in dynamic SMG condition exhibited looser structures and better cell viabilities with flat and regular surfaces. Therefore, the KSR media promotes spherical formation by hADSCs, which showed polar growth, fusion, and injectability in vitro. The dynamic SMG culture enhances the formation of a looser structure and better cell viabilities for hADSC spheroids.

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Keywords

hADSCs; EP tube; Spheroids; Microgravity

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