Expansion and hepatic differentiation of rat multipotent adult progenitor cells in microcarrier suspension culture

Abstract

Many potential applications of stem cells require large quantities of cells, especially those involving large organs such as the liver. For such applications, a scalable reactor system is desirable to ensure a reliable supply of sufficient quantities of differentiation competent or differentiated cells. We employed a microcarrier culture system for the expansion of undifferentiated rat multipotent adult progenitor cells (rMAPC) as well as for directed differentiation of these cells to hepatocyte-like cells. During the 4-day expansion culture, cell concentration increased by 85-fold while expression level of pluripotency markers were maintained, as well as the MAPC differentiation potential. Directed differentiation into hepatocyte-like cells on the microcarriers themselves gave comparable results as observed with cells cultured in static cultures. The cells expressed several mature hepatocyte-lineage genes and asialoglycoprotein receptor-1 (ASGPR-1) surface protein, and secreted albumin and urea. Microcarrier culture thus offers the potential of large-scale expansion and differentiation of stem cells in a more controlled bioreactor environment.

Bistability in Glycolysis Pathway as a Physiological Switch in Energy Metabolism

“…HeLa cells were inoculated at 2×10 5 cells/mL on Day 0. The procedure for microcarrier culture has been described previously [60]. On Day 5, cells on microcarriers were washed and re-suspended at 1×10 5 cells/mL in DMEM medium containing either 0.6 mM (LG) or 25 mM (HG) of glucose.…”

Titanium-Enriched Hydroxyapatite–Gelatin Scaffolds with Osteogenically Differentiated Progenitor Cell Aggregates for Calvaria Bone Regeneration

“…14,15 They also have high levels of the Oct4 transcription factor, a high growth rate, and are able to be grown as 3D aggregates. 16,17 MAPCs are also capable of forming the three germ lineages, including the mesoderm that gives rise to bone tissue when dexamethasone is supplemented. 18,19 Recent studies demonstrate that clinical grade human MAPCs (Multistem Ò ) offer clinical relevance for the treatment of autoimmune disorders and cardiac infarction (ClinicalTrials.gov NCT00677859, NCT00677222); however, no in vivo applicability studies for bone regeneration have been reported.…”

The road to regenerative liver therapies: The triumphs, trials and tribulations

“…Any clinical cellular therapy or bioartificial liver applications will likely employ at least 5–10% of that cell number in order to to provide sufficient functional activities, or to shorten the time frame of regeneration. With a typical bioreactor, cultivating cells as aggregates [85] or on microcarriers [86], a cell concentration of at least 10 9 cell/L is easily achievable. Assuming a manufacturing process of preparing 100 doses per batch, the size of the reactor is in the range of 1000 L. Reactors with thousands of liters in volume are commonly seen in the production of viral vaccines and protein biologics [87].…”