Mesenchymal Stem Cell Isolation from Human Umbilical Cord Tissue: Understanding and Minimizing Variability in Cell Yield for Process Optimization

Abstract

Human tissue banks are a potential source of cellular material for the nascent cell-based therapy industry; umbilical cord (UC) tissue is increasingly privately banked in such facilities as a source of mesenchymal stem cells for future therapeutic use. However, early handling of UC tissue is relatively uncontrolled due to the clinical demands of the birth environment and subsequent transport logistics. It is therefore necessary to develop extraction methods that are robust to real-world operating conditions, rather than idealized operation. Cell yield, growth, and differentiation potential of UC tissue extracted cells was analyzed from tissue processed by explant and enzymatic digestion. Variability of cell yield extracted with the digestion method was significantly greater than with the explant method. This was primarily due to location within the cord tissue (higher yield from placental end) and time delay before tissue processing (substantially reduced yield with time). In contrast, extraction of cells by explant culture was more robust to these processing variables. All cells isolated showed comparable proliferative and differentiation functionality. In conclusion, given the challenge of tightly controlled operating conditions associated with isolation and shipping of UC tissue to banking facilities, explant extraction of cells offers a more robust and lower-variability extraction method than enzymatic digestion.

Isolation, cultivation, and characterization of human mesenchymal stem cells

“…Only few studies have directly compared explant UC cultures to enzymatic isolation strategies. For example, Buyl et al and Iftimia‐Mander et al performed an explant culture and enzymatic isolation using 0.05% (m/v) collagenase type II solution or 0.075% Collagenase type I solution respectively of uc‐MSC. Both reported similar cell morphology and comparable doubling times for the different procedures.…”

Therapeutic efficacy of neural stem cells originating from umbilical cord-derived mesenchymal stem cells in diabetic retinopathy

“…MSCs are limited to bone marrow and several types of connective tissue in adults 3 . They are capable of cloning to form adherent fibroblastic cells that express the unique properties of the cell surface phenotype.…”

Umbilical Cord Blood NOS1 as a Potential Biomarker of Neonatal Encephalopathy

“…Umbilical cords were obtained from healthy full-term births within 4 h. Methods previously described by Thomas et al (27); briefly, in a sterile cell culture hood, umbilical cord was cut into 1-mm length thick slices. For normoxia, specimens were cultured in a conventional incubator preset at 37°C in a humidified atmosphere, containing 5% CO 2 .…”