pnc-Si bioreactor for red blood cell culture!

Hi all, this is Henry.

Leeanna Hyacinth, an undergraduate student from the McNair program, has recently completed a joint project between the Waugh, Palis, and McGrath Lab to create a bioreactor to amplify the number of mature red blood cells.

Background:

The Palis lab has discovered a method to amplify erythroid precursor cells (from 1 to 10^30 cells) and start their differentiation into red blood cells.  However, many of the differentiated cells are not fully matured red blood cells.  By matured, we mean the attainment of enucleation and membrane stability.

The Waugh lab have showed through their studies that during late-stage erythropoiesis, there is a transient period of membrane instability.   However, it’s been demonstrated previously that the injection of red blood cell precursors into mice bone marrow have successively produced matured red blood cell.  It is possible that there are mechanisms  in the bone marrow, such as the close packing of cells, that stabilizes membranes against surface loss during maturation .

Hypothesis: The close compaction of erythroid precusors in the differentiation media will promote their maturation into red blood cells.

Problem: How does one closely compact the erythroid precursor cells?

Possible solution: A bioreactor with pnc-Si as an integral component.  Fluid flow was driven with cell-containing media through the pnc-Si.  The cells are retained and closely compacted onto the pnc-Si because they are much larger than the pores on the membrane.

Significance: If we are able to create large quantity of mature red blood cells from the erythroid precursor cells, it will solve the blood shortage problem in various ways.  No longer is there a need for large number of healthy donors.  Any type of red blood cells (A, B, AB, O, etc.) can be created.  There will also be less complications due to infectious diseases.

Methods and Results:

Siphoning of RBC precursors onto pnc-Si.  There is a reperfusion of cells at frame 53.  (The total duration of imaging is 30 min): Leeanna_Jul_22_2009 syphoning (compressed)

Reverse siphoning of RBC  precrusors to release the cell from the pnc-Si.  (The total duration of imaging is 10 min):

Leeanna_Jul_22_2009 reversal for cell release (compressed)

Schematics of the RBC bioreactor

Comparison of cells (normal culture vs culture in pnc-Si RBC bioreactor):

Legend: Mix of bone marrow and E12.5 precursor red cell in culture after 72 hours in differentiation medium. A: control shows some hemoglobin dense cells. B, C: cells from bioreactor.