In vitro and clinical evaluation of umbilical cord-derived mesenchymal stromal cell-conditioned media for hair regeneration.

BACKGROUND: The field of regenerative medicine may present a non-drug, non-steroid, and non-invasive alternative toward addressing male and female pattern hair loss, a global concern. OBJECTIVE: The aim was to carry out the in vitro and in vivo safety and efficacy evaluation of human umbilical cord-derived mesenchymal stromal cell-conditioned media (MSC-CM) for hair regeneration. METHODS: Various in vitro parameters were used to estimate the consistency across various batches of MSC-CM. Total protein content was measured by the Biuret method and antioxidant activity by the 2,2-diphenyl-1-picryl hydrazyl (DPPH) assay. Fourier transform infrared spectroscopy (FTIR) analysis was used to determine spectral signatures and biocompatibility was carried out by the Neutral Red Uptake (NRU) and Sulforhodamine B (SRB) assays. In vivo safety and efficacy was evaluated in an experimental pilot study on 15 volunteers. RESULTS: The in vitro results confirmed stability in the protein content (7 mg/ml), antioxidant activity (49.50%), and FTIR fingerprints of the MSC-CM. In the biocompatibility experiments by both SRB and NRU methods, no IC50 value could be derived at 100% concentration indicating safety at the cellular level. The in vivo results indicated safety with no side effects or adverse reactions, while 86.6% of the subjects experienced a positive effect of hair regeneration. CONCLUSION: MSC-CM comprises a rich cocktail of physiologically balanced growth factors, cytokines, and beneficial proteins which may explain the bioactivity and mechanism of action in hair regrowth. This may indicate a biocompatible, gentle, and safe regenerative approach to address hair loss.

Xeno-Free Human Wharton's Jelly Mesenchymal Stromal Cells Maintain Their Characteristic Properties after Long-Term Cryopreservation.

OBJECTIVE: The past decade has witnessed a rapid growth in harnessing the potential of adult stem cells for regenerative medicine. An investigational new drug (IND) or a regenerative medicine advanced therapy (RMAT) product must fulfil many requirements, such as stability studies, after cryopreservation. Such studies are important to ascertain the utility of off-the-shelf allogeneic cells for clinical applications. The present work describes a complete characterisation of xenofree human Wharton's Jelly mesenchymal stromal cells (hWJ-MSCs) before and up to 28 months post-cryopreservation. MATERIALS AND METHODS: In this experimental study, culture methods that involved plasma derived human serum and recombinant trypsin were used to develop clinical grade cells. Complete cell characterisation involved flow cytometry studies for viability, positive and negative markers, colony forming unit (CFU) potential, population doubling time (PDT), soft agar assay to evaluate in vitro tumourigenicity, karyotype analysis and differentiation studies which were performed before and at 6, 12, 18 and 28 months post-cryopreservation. RESULTS: Our data showed consistency in the flow cytometry, CFU assay, PDT, soft agar assay, karyotyping and differentiation studies. CONCLUSION: Using our protocols for extended xeno-free culture and cryopreservation of hWJ-MSCs, we could establish the shelf life of the cell-based product for up to 28 months.

Evaluation of Potential Application of Wharton's Jelly-Derived Human Mesenchymal Stromal Cells and its Conditioned Media for Dermal Regeneration using Rat Wound Healing Model.

Mesenchymal stromal cells and the derived conditioned media represent an area of tremendous medical interest and, among other clinical applications, are currently being extensively explored for wound healing. The aim of this study was to comparatively evaluate the wound healing potential of xeno-free human umbilical cord-derived mesenchymal stromal cells (MSCs) and the conditioned media (CM) in a full-thickness excision wound model in rats. The evaluation parameters included rate of wound healing, serum cytokine analyses, collagen content, histopathology, and hyperspectral imaging as an independent qualitative and quantitative tool. Both the cell-based and cell-free approaches scored better in lower inflammation, as evidenced in lower IL-10 and stable IL-6 levels, and improved rate of wound healing (p < 0.0001). More importantly, no adverse reaction or rejection was observed although human MSCs and CM were used in a xenogeneic model. The presence of hFGF, hHGF, hGCSF, hIL-1Ra, hVEGF, and hIL-6 in the secretome may elucidate the regenerative potential of the xeno-free cell-based and cell-free approaches which have translational value for advanced wound care. The results revealed the therapeutic potential of both the cell-based and cell-free approaches for wound healing.