One of the primary obstacles that stands with respect to using human hematopoietic stem cells (hHSCs) to deal with various diseases could be the difficulty growing them in culture—they quickly die or differentiate into other cell types. Several experiments that demonstrate the successful usage of fat cells included in a feeder layer to aid prolonged increase of hHSCs in culture is reported in an article in BioResearch Open Access, a bimonthly peer-reviewed open access journal from Mary Ann Liebert, Inc., publishers. The content can be found within the BioResearch Open Access website.
Inside the article “Extending Human Hematopoietic Stem Cell Survival In Vitro with Adipocytes” Dean Liang Glettig and David Kaplan, Tufts University, Medford, MA included adipocytes (fat cells) in varying amounts and locations within the feeder layers of hHSCs being grown inside the laboratory. They varied the concentrations of various cell types including adipocytes inside feeder layer, comparing different degrees of adipocytes, and evaluated the effect of direct cell-to-cell contact between the hHSCs along with the adipocytes inside feeder layer on the survival rate from the hHSCs.
“The opportunity to prolong hHSC culture in vitro not simply benefits basic stem cell research, it is additionally an important step towards developing advanced cell therapies for future clinical use,” says BioResearch Open Access Editor Jane Taylor, PhD, MRC Centre for Regenerative Medicine, University of Edinburgh, Scotland.
The abstract of this article is as following.
Human hematopoietic stem cells (hHSCs) cannot be maintained in vitro for extended time periods because they rapidly differentiate or die. To extend in vitro culture time, researchers have made attempts to use human mesenchymal stem cells (hMSCs) to create feeder layers that mimic the stem cell niche. We have conducted an array of experiments including adipocytes in these feeder layers that inhibit hHSC differentiation and by that prolong stem cell survival in vitro. The amount of CD34+ cells was quantified using flow cytometry. In a first experiment, feeder layers of undifferentiated hMSCs were compared with feeder layers differentiated toward osteoblasts or adipocytes using minimal medium, showing the highest survival rate where adipocytes were included. The same conclusion was drawn in a second experiment in comparing hMSCs with adipogenic feeder cells, using a culture medium supplemented with a cocktail of hHSC growth factors. In a third experiment, it was shown that direct cell–cell contact is necessary for the supportive effect of the feeder layers. In a fourth and fifth experiment the amount of adipocytes in the feeder layers were varied, and in all experiments a higher amount of adipocytes in the feeder layers showed a less rapid decay of CD34+ cells at later time points. We therefore concluded that adipocytes assist in suppressing hHSC differentiation and aid in prolonging their survival in vitro.
Read full article here.
Source: Dean Liang Glettig and David L. Kaplan. BioResearch Open Access. “Extending Human Hematopoietic Stem Cell Survival In Vitro with Adipocytes“. doi:10.1089/biores.2013.0006.
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