A recent article published in the International Journal of Molecular Sciences utilized CHPs to verify the health of collagen in their MSC-ECM complexes. The authors developed a clumped-MSC/ECM (C-MSC) complex under serum-free conditions that allows bone regeneration upon implantation without the use of an artificial scaffold. They grew the C-MSC and compared a cellularized vs decellularized C-MSCs for their ability to promote osteogenesis after implantation. F-CHPs were used to verify that there was no damage to the ECM after decellularization procedures. In either case, there was no damage seen to the collagen within the ECM complex. You only see F-CHP signal in the positive control.

Abstract: Three-dimensional clumps of mesenchymal stem cell (MSC)/extracellular matrix (ECM) complexes (C-MSCs) consist of cells and self-produced ECM. We demonstrated previously that C-MSCs can be transplanted into bone defect regions with no artificial scaffold to induce bone regeneration. To apply C-MSCs in a clinical setting as a reliable bone regenerative therapy, the present study aimed to generate C-MSCs in xeno-free/serum-free conditions that can exert successful bone regenerative properties and to monitor interactions between grafted cells and host cells during bone healing processes. Human bone marrow-derived MSCs were cultured in xeno-free/serum-free medium. To obtain C-MSCs, confluent cells that had formed on the cellular sheet were scratched using a micropipette tip and then torn off. The sheet was rolled to make a round clump of cells. Then, C-MSCs were transplanted into an immunodeficient mouse calvarial defect model. Transplantation of C-MSCs induced bone regeneration in a time-dependent manner. Immunofluorescence staining showed that both donor human cells and host mice cells contributed to bone reconstruction. Decellularized C-MSCs implantation failed to induce bone regeneration, even though the host mice cells can infiltrate into the defect area. These findings suggested that C-MSCs generated in xeno-free/serum-free conditions can induce bone regeneration via direct and indirect osteogenesis.

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