Biotechnol Lett. 2014 Jul;36(7):1529-37.

Maintenance of human adipose derived stem cell (hASC) differentiation capabilities using a 3D culture.

Lin CY, Huang CH, Wu YK, Cheng NC, Yu J.

Contact: Jiashing Yu. Email: jiayu@ntu.edu.tw

 

Abstract

In this study, 3D culture system for human adipose-derived stem cell (hASC) using a BioLevitator as the bioreactor for microcarrier-based cultures was established. During the culturing period, hASCs preferred to grow in crevices between microcarriers and a high viability was maintained even when reaching confluency. Adipogenic or osteogenic differential medium was used to induce hASCs and differential potentials of these cells were compared between 2D and 3D environments via RT-PCR and staining quantifications. CEBP/α gene expression was significant higher in 3D condition at day 21 (P < 0.05). Staining quantification indicates that cells cultured in 3D condition have significant better differentiation potential from day 14 to 21 for both adipogenic and osteogenic lineages (P < 0.01).

PMID: 24658740

 

Supplements:

Adipose-derived stem cells (ASCs) are stem cells with multipotency and differential ability toward the osteogenic, adipogenic, myogenic and chondrogenic lineages. Compared to the relatively non-abundant and difficult-to-access bone marrow and embryonic stem cells, ASCs can be easily isolated from adipose tissue with minimal donor discomfort. ASCs meet the basic criteria for an ideal stem cell sources in regenerative medicinal applications: abundant quantities, minimal invasive harvest procedure, multiple cell differential lineage pathways, safe and effective transplantation to either autologous or allogeneic host. The similar characteristics between ASCs and bone marrow mesenchymal stem cells (BMMSCs) indicate that ASCs might have the potential to be a better cell source in stem cell research and regenerative medicine study.

 

3D culture WBF

Fig 1. Illustration of 3D culture

 

Differentiation of 2D and 3D culture WBF

Fig 2. Differentiation of 2D and 3D culture.

 

With the possible benefits of 3D culture, we establishws a 3D culture system for different cell types using BioLevitatorTM as the bioreactor for microcarrier-based culture, and further compare the differential potential of hASC in 2D/3D environment. Unlike general stirring bioreactor with impeller, BioLevitatorTM provide a thorough mixing environment via the blades at the bottom of the culture tube. Moreover, tunable parameters of the system include agitation time, rotation direction, speed and duration. With these multiple parameters, even attachment on microcarriers within a short period of time can be achieved by tuning agitation and rotating period. Relatively higher rotation speed in culture stage prevents the cell-microcarriers from aggregating. Magnetic microcarrier GEM and cytodex3 are used as the microcarriers for our 3D culture.

With this 3D microcarrier culture system, we successfully expanded the cell population in a short period of time. By adipogenic and osteogenic differential medium induction of human ASCs previously cultured in 2D or 3D culture environment, we would test whether adipose-derived stem cell can improve their differential capability after introducing into a 3D culture environment. We look forward to further investigator of the harvested cells transplanted in vivo as well as detailed molecular biology examination of the cells obtained with the demonstrate d method.

 

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