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In vitro cultured plant cells, in particular the tobacco BY-2 cell, have demonstrated their potential to provide a promising bioproduction platform for therapeutic proteins by integrating the merits of whole-plant cultivation systems with those of microbial and mammalian cell cultures. Over the past three decades, substantial progress has been made in improving the plant cell culture system, resulting in a few commercial success cases, such as taliglucerase alfa (Elelyso®), the first FDA-approved recombinant pharmaceutical protein derived from plant cells. However, compared to the major expression hosts (bacteria, yeast, and mammalian cells), plant cells are still largely underutilized, mainly due to low productivity and non-human glycosylation. Modern molecular biology tools, in particular RNAi and the latest genome editing technology CRISPR/Cas9, have been used to modulate the genome of plant cells to create new cell lines that exhibit desired “traits” for producing therapeutic proteins. This review highlights the recent advances in cellular engineering of plant cells towards improved recombinant protein production, including creating cell lines with deficient protease levels or humanized glycosylation, and considers potential development in the future.

Ex vivo generation of red blood cells (RBCs) from hematopoietic stem cells (HSCs) used for blood transfusion represents one of the focuses in current regenerative medicine. However, massive production of HSCs-based RBCs requires a significant quantity of erythropoietic growth factors, making manufacturing at large scale cost prohibitive. Plant cell culture is proposed to be a promising bioproduction platform for functional human proteins in a safe and cost-efficient manner. This study exploited a proprietary technology, named HypGP engineering technology, for high-yield production of one of the key erythropoietic growth factors--stem cell factor (SCF)--in plant cell culture. Specifically, a designer hydroxyproline (Hyp)-O-glycosylated peptide (HypGP) comprised of 20 tandem repeats of the “Ser-Pro” motif, or (SP)(20,) was engineered at either the N-terminus or C-terminus of SCF in tobacco BY-2 cells. The (SP)(20) tag dramatically increased the secreted yields of SCF up to 2.5 μg/ml. The (SP)(20)-tagged SCF showed bioactivity in promoting the proliferation of the TF-1 cell line, although the SCF-(SP)(20) was 8.4-fold more potent than the (SP)(20)-SCF. Both the (SP)(20)-SCF and SCF-(SP)(20) exhibited desired function in stimulating the expansion and differentiation of human umbilical cord blood CD34(+) cells towards RBCs.