Creation of GMP-Compliant iPSCs From Banked Umbilical Cord Blood

Abstract

Many clinical trials are in progress using cells derived from induced pluripotent stem cells (iPSC) for immunotherapies and regenerative medicine. The success of these new therapies is underpinned by the quality of the cell population used to create the iPSC lines, along with the creation of iPSCs in a fully Good Manufacturing Practice (GMP)-compliant environment such that they can be used safely and effectively in the clinical setting. Umbilical cord blood (CB) from public cord blood banks is an excellent source of starting material for creation of iPSCs. All CB units are manufactured under GMP-conditions, have been screened for infectious diseases, with known family and medical history of the donor. Furthermore, the HLA tissue typing is known, thereby allowing identification of CB units with homozygous HLA haplotypes. CB cells are naïve with less exposure to environmental insults and iPSC can be generated with high efficiency. We describe a protocol that can be adopted by those seeking to create clinical-grade iPSC from banked CB. This protocol uses a small volume of thawed CB buffy to first undergo ex-vivo expansion towards erythroid progenitor cells, which are then used for reprogramming using the CytoTune™-iPS 2.0 Sendai Reprogramming Kit. Resultant iPSC lines are tested to confirm pluripotency, genomic integrity, and stability. Cells are maintained in a feeder-free, xeno-free environment, using fully defined, commercially available reagents. Adoption of this protocol, with heed given to tips provided, allows efficient and robust creation of clinical-grade iPSC cell lines from small volumes of cryopreserved CB.

Perspectives for the Use of Umbilical Cord Blood in Transplantation and Beyond: Initiatives for an Advanced and Sustainable Public Banking Program in Greece

“…The high rejection rate of the donated UCB units due to the strict acceptance criteria for HSCT purposes, in association with the high application of the haploidentical HSCT, have resulted in a significant limitation of the UCB use in the clinic, with a consequent impact on the economic sustainability of the public UCB banks. The perspective of recycling the inappropriate for HSCT units or using the high-quality excess units for isolation and expansion of immune and stem cell populations for cell engineering/reprogramming processes and for production of cell-derivatives for research and clinical applications, extends the therapeutic value of the UCB and increases the sustainability potential of the UCB banks [18,[21][22][23]35,[38][39][40][41][42]52,65,68,70,75,[78][79][80][81][82][83][84][85][86][97][98][99][100]. Novel applications in adoptive immunotherapy, regenerative medicine and specialized transfusion are anticipated not only to maximize the use of the UCB but also to minimize the financial deficit of the public UCB banks through reimbursement procedures.…”

A comprehensive human leukocyte antigen analysis of 36 782 cord blood units stored in the Australian Public Cord Blood Banking Network

“…This analysis has identified common tissue types and haplotypes that are valuable research resources due to their potential wide scope of utility in the Australian and global population. It has also identified within each bank the CBU with common homozygous haplotypes that may be of benefit for CB-derived induced pluripotent stem cell haplobanking strategies [17]. Future analyses may focus on highlighting differences between the AusCord CBB inventory and the rest of the international inventory.…”

Induced pluripotent stem cells in companion animals: how can we move the field forward?

“…Umbilical cord blood banks are also a useful resource for iPSC reprogramming (50,51). The differentiation status of the cell can influence the reprogramming efficiency, as the epigenomic state needs to be reset to a pluripotent state.…”

“…Indeed, waste or discarded tissues, such as foreskin fibroblasts ( 47 ), periodontal tissue ( 48 ), or renal cells from urine ( 49 ) have also been taken used. Umbilical cord blood banks are also a useful resource for iPSC reprogramming ( 50 , 51 ).…”