Towards the standardization of methods of tissue processing for the isolation of mesenchymal stromal cells for clinical use

Abstract

Multipotent mesenchymal stromal cells (MSCs) are currently the most extensively studied type of adult stem cells in advanced stages of development in the field of regenerative medicine. The biological properties of MSCs have generated great hope for their therapeutic use in degenerative and autoimmune conditions that, at present, lack effective treatment options. Over the last decades, MSCs have been typically obtained from adult bone marrow, but the extraction process is highly invasive and the quality and numbers of isolated cells is drastically influenced by patient age, medication and associated comorbidities. Therefore, there is currently an open discussion on the convenience of allogeneic over autologous treatments, despite potential disadvantages such as rejection by the host. This shift to the allogeneic setting entails the need for high production of MSCs to ensure availability of sufficient cell numbers for transplantation, and therefore making the search for alternative tissue sources of highly proliferative MSC cultures with low levels of senescence occurrence, which is one of the greatest current challenges in the scale up of therapeutic cell bioprocessing. Herein we (i) present the main isolation protocols of MSCs from bone marrow, adipose tissue and Wharton’s jelly of the umbilical cord; and (ii) compare their qualities from a bioprocess standpoint, addressing both quality and regulatory aspects, in view of their anticipated clinical use.

Preclinical Development of a Therapy for Chronic Traumatic Spinal Cord Injury in Rats Using Human Wharton’s Jelly Mesenchymal Stromal Cells: Proof of Concept and Regulatory Compliance

“…Clinical grade ex vivo expanded MSC were derived from the Wharton’s jelly of human umbilical cords following current Good Manufacturing Practice (GMP)-compliant methods reported in detail elsewhere [ 16 , 17 ]. Cells were expanded in vitro up to sufficient numbers (≤5 passages) for further testing by using Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco) containing 2 mM glutamine supplemented with 10% human serum (hSer) B [ 9 , 16 ].…”

Optimized reagents for immunopotency assays on mesenchymal stromal cells for clinical use

“…PBMCs were used individually or pooled, fresh or cryopreserved, according to the experimental design. MSC were derived from the Wharton’s Jelly (WJ) of donated umbilical cords following GMP-compliant procedures reported elsewhere [ 29 , 30 ], with appropriate donor informed consent. After the isolation step, cells were replated by seeding T cell culture flasks at 1.5–3 × 10 3 cell/cm 2 .…”

Bioactive ECM-Loaded SIS Generated by the Optimized Decellularization Process Exhibits Skin Wound Healing Ability in Type I Diabetic Rats

“…3,4 Clinically, the surgical management of diabetic ulcers often involves use of skin flaps and/or full-thickness skin grafts, which are risky for patients since they require a secondary surgery, and the graft is difficult to survive. 5,6 With the rapid development of tissue engineering and regeneration medicine, human mesenchymal stem cells (MSCs) derived from bone marrow, adipose tissue, and umbilical cord 7 have been used as seed cells to promote diabetic wound healing. 8 However, the acquisition of MSCs often requires invasive surgery.…”