TY - JOUR

T1 - Colonization of collagen scaffolds by adipocytes derived from mesenchymal stem cells of the common marmoset monkey

AU - Bernemann, Inga

AU - Mueller, Thomas

AU - Blasczyk, Rainer

AU - Glasmacher, Birgit

AU - Hofmann, Nicola

N1 - Funding Information: We thank Resorba®, Germany, for the provision of the collagen and the Centre for Reproductive Medicine and Andrology Muenster (CeRA) for donation of the marmoset bone marrow. Thanks to A. Deiwick and C. Marx from IMP, Leibniz Universität Hannover, K. Egler from the MH Hannover for their outstanding technical support and W. Hake for his excellent photographic work ( Fig. 1 A). This work is supported by funding from the Deutsche Forschungsgemeinschaft (DFG, German research Foundation) for the Cluster of Excellence REBIRTH (from Regenerative Biology to Reconstructive Therapy) (EXC 62/1).

PY - 2011/7/29

Y1 - 2011/7/29

N2 - In regenerative medicine, human cell replacement therapy offers great potential, especially by cell types differentiated from immunologically and ethically unproblematic mesenchymal stem cells (MSCs). In terms of an appropriate carrier material, collagen scaffolds with homogeneous pore size of 65 μm were optimal for cell seeding and cultivating. However, before clinical application and transplantation of MSC-derived cells in scaffolds, the safety and efficiency, but also possible interference in differentiation due to the material must be preclinically tested. The common marmoset monkey (Callithrix jacchus) is a preferable non-human primate animal model for this aim due to its genetic and physiological similarities to the human.Marmoset bone marrow-derived MSCs were successfully isolated, cultured and differentiated in suspension into adipogenic, osteogenic and chondrogenic lineages by defined factors. The differentiation capability could be determined by FACS. Specific marker genes for all three cell types could be detected by RT-PCR. Furthermore, MSCs seeded on collagen I scaffolds differentiated in adipogenic lineage showed after 28. days of differentiation high cell viability and homogenous distribution on the material which was validated by calcein AM and EthD staining. As proof of adipogenic cells, the intracellular lipid vesicles in the cells were stained with Oil Red O. The generation of fat vacuoles was visibly extensive distinguishable and furthermore determined on the molecular level by expression of specific marker genes. The results of the study proved both the differential potential of marmoset MSCs in adipogenic, osteogenic and chondrogenic lineages and the suitability of collagen scaffolds as carrier material undisturbing differentiation of primate mesenchymal stem cells.

AB - In regenerative medicine, human cell replacement therapy offers great potential, especially by cell types differentiated from immunologically and ethically unproblematic mesenchymal stem cells (MSCs). In terms of an appropriate carrier material, collagen scaffolds with homogeneous pore size of 65 μm were optimal for cell seeding and cultivating. However, before clinical application and transplantation of MSC-derived cells in scaffolds, the safety and efficiency, but also possible interference in differentiation due to the material must be preclinically tested. The common marmoset monkey (Callithrix jacchus) is a preferable non-human primate animal model for this aim due to its genetic and physiological similarities to the human.Marmoset bone marrow-derived MSCs were successfully isolated, cultured and differentiated in suspension into adipogenic, osteogenic and chondrogenic lineages by defined factors. The differentiation capability could be determined by FACS. Specific marker genes for all three cell types could be detected by RT-PCR. Furthermore, MSCs seeded on collagen I scaffolds differentiated in adipogenic lineage showed after 28. days of differentiation high cell viability and homogenous distribution on the material which was validated by calcein AM and EthD staining. As proof of adipogenic cells, the intracellular lipid vesicles in the cells were stained with Oil Red O. The generation of fat vacuoles was visibly extensive distinguishable and furthermore determined on the molecular level by expression of specific marker genes. The results of the study proved both the differential potential of marmoset MSCs in adipogenic, osteogenic and chondrogenic lineages and the suitability of collagen scaffolds as carrier material undisturbing differentiation of primate mesenchymal stem cells.

KW - Adipogenic differentiation

KW - Collagen scaffolds

KW - Common marmoset monkey

KW - MSCs

KW - Non-human primate

UR - http://www.scopus.com/inward/record.url?scp=79960836886&partnerID=8YFLogxK

U2 - 10.1016/j.bbrc.2011.06.134

DO - 10.1016/j.bbrc.2011.06.134

M3 - Article

C2 - 21726528

AN - SCOPUS:79960836886

VL - 411

SP - 317

EP - 322

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 2

ER -