Details

Autor(en) / Beteiligte

• Nienow, Alvin W.
• Rafiq, Qasim A.
• Coopman, Karen
• Hewitt, Christopher J.

Titel

A potentially scalable method for the harvesting of hMSCs from microcarriers

Ist Teil von

• Biochemical engineering journal, 2014-04, Vol.85, p.79-88

Ort / Verlag

Amsterdam: Elsevier B.V

Erscheinungsjahr

2014

Link zum Volltext

Quelle

Elsevier ScienceDirect Journals Complete

Beschreibungen/Notizen

• •A scalable method based on agitation theory for harvesting hMSCs from microcarriers.•Harvesting involves cell detachment from microcarriers followed by separation step.•A combined short burst of intense agitation plus enzyme detaches cells in ~7min.•After detachment, cells are not be damaged being smaller than Kolmogorov microscale.•After harvest, cells retain all their usual quality attributes. The use of hMSCs for allogeneic therapies requiring lot sizes of billions of cells will necessitate large-scale culture techniques such as the expansion of cells on microcarriers in bioreactors. Whilst much research investigating hMSC culture on microcarriers has focused on growth, much less involves their harvesting for passaging or as a step towards cryopreservation and storage. A successful new harvesting method has recently been outlined for cells grown on SoloHill microcarriers in a 5L bioreactor [1]. Here, this new method is set out in detail, harvesting being defined as a two-step process involving cell ‘detachment’ from the microcarriers’ surface followed by the ‘separation’ of the two entities. The new detachment method is based on theoretical concepts originally developed for secondary nucleation due to agitation. Based on this theory, it is suggested that a short period (here 7min) of intense agitation in the presence of a suitable enzyme should detach the cells from the relatively large microcarriers. In addition, once detached, the cells should not be damaged because they are smaller than the Kolmogorov microscale. Detachment was then successfully achieved for hMSCs from two different donors using microcarrier/cell suspensions up to 100mL in a spinner flask. In both cases, harvesting was completed by separating cells from microcarriers using a Steriflip® vacuum filter. The overall harvesting efficiency was >95% and after harvesting, the cells maintained all the attributes expected of hMSC cells. The underlying theoretical concepts suggest that the method is scalable and this aspect is discussed too.