Table 1.
Progression in the field of ex vivo RBCs expansion during last two decades.
| S. no. | Cell source and culture time | Growth factors | Co-culture | Expansion | Units of blood | Enucleation rate | Animal or human serum/plasma | Key points | References |
|---|---|---|---|---|---|---|---|---|---|
| 1 | CB CD34+ 21 days | FLT-3, SCF, TPO, EPO, IGF-1 | NO | NA | NA | 4% in vitro | No | Basis for transfusion potential of ex vivo cultured RBCs in animal model | Neildez-Nguyen et al., 2002 |
| 99% in vivo | |||||||||
| 2 | Cord blood 21 days | SCF, IL-3, EPO hydrocortisone | Murine stromal mMS-5 | 1.95 × 106 | 4.6/CB | 95% | No | First in vitro production of RBCs | Giarratana et al., 2005 |
| 3 | Cord blood 60 days | EPO, SCF, IGF-1 dex and lipid mix | No | 109 | -NA | −100% | Yes | Prolonged expansion protocol up to 60 days for adult globin switching | Leberbauer et al., 2005 |
| 4 | Cord blood 20 days | SCF, IL-3, EPO VEGF, IGF-2 | No | 7.2 × 105 | 104/CB | 77.5% | Yes | Low expansion and enucleation rates | Miharada et al., 2006 |
| 5 | Cord blood 21 days | SCF, IL-3, EPO, TPO, Flt-3 | Human MSCs | 8 × 103 | 0.02/CB | 64% | No | Less allogenicity, lower expansion, and enucleation rates, replaced BM derived feeder cells with UCB derived cells | Baek et al., 2008 |
| 6 | Cord blood 38 days | SCF+Flt-3+TPO IL-3+EPO | hTERT+ Macrophages | 3.5 × 106 | 8.8/CB | 100% | Yes | Impossible to scale up | Fujimi et al., 2008 |
| 7 | hESCs 59 days | Hydrocortisone, IL3, BMP-4, SCF, EPO, IGF-1 | hMSCs, mMS-5 | 4 × 107 cells | NA | (6.5%, starting from CD34+ cells) | Yes | Globin switching | Qiu et al., 2008 |
| FH-B-hTERT | |||||||||
| 8 | hESCs 42 days | SCF, Epo, BMP-4, VEGF, bFGF, TPO, FLT3 L | MEFs, OP9 | 1010–1011 cells/6-well plate of hESCs | NA | 10–65% | No | Functional oxygen carrying capacity of ESCs derived RBCs | Lu et al., 2008 |
| 10 | hiPSCs,(MR90, FD-136) | SCF,TPO, FLT3-L, TPO, BMP-4; VEGF- IL-3, IL-6 | hIPSCs | NA | 4–10% | No | First time complete differentiation of hiPSCs cells into definitive erythrocytes capable of maturation up to enucleated RBCs (fetal hemoglobin in a functional tetrameric form) | Lapillonne et al., 2010 | |
| hESCs(H1I) 46 days | 4.4 × 108 | 52–66% | |||||||
| hESCs 35 × 108 | |||||||||
| 11 | Cord blood 33 days | SCF, IL-3, EPO hydrocortisone | No | 2.25 × 108 | −500 units/UCB | >90% | No | First serum-free culture, first demonstration of RBC culture in a large-scale bioreactor | Timmins et al., 2011 |
| 12 | Cord blood 18 days | SCF, EPO hydrocortisone | No | 4.3 × 107 | 75/CB | 70% | Yes | Best yield to date | Giarratana et al., 2011 |
| 13 | hiPSCs 59 days | Hydrocortisone, IL-3 BMP-4, Flt3L, SCF, EPO, IGF-1 | hMSCs/Matrigel. | 0.5–8 × 106 | NA | NA | Yes | Production of large number of erythroid cells with embryonic and fetal-like characteristics regardless of the age of donor tissue | Chang et al., 2011 |
| hTERT immortalized fetal liver cells | |||||||||
| 14 | hESCs hiPSCs 60–125 days | Iron saturated transferrin, dexamethasone, insulin, SCF, EPO, TPO, IL-3, IL-6 | MEFs, mOP9 mMS-5 | 2 × 105 ESCs | NA | 2–10% | Yes | RBCs production from transgenic and transgene-free iPSCs using the OP9 coculture method with efficiency comparable to hESCs | Dias et al., 2011 |
| 15 | hiPSCs 52 days | holo-human transferrin recombinant human insulin heparin, and 5% human plasma SCF, TPO, FLT3 ligand, BMP4, VEGF- n-3, IL-Epo | MEFs | 15–28.3 × 108 | NA | 20–26% RBC and 74–80% orthochromatic erythroblasts | 5–10% Hu plasma | First time in a normal and a pathological erythropoietic differentiation models that hiPSC are intrinsically able to mature into adult hemoglobin synthesizing cells | Kobari et al., 2012 |