Table 1.

Preparation of iPSCs for personalized medicine

iPSC Preparation Stage	Techniques	Features	Outcomes	References
Reprogramming Techniques	Lentiviruses	iPSCs produced from adult fibroblast	Treatment with valproic acid increased cell proliferation	[21]
	Lentiviruses	iPSCs produced from mouse tail-tip fibroblast	Porphyra 334 increased the effectiveness of cell reprogramming	[22]
	Sendai viruses	iPSCs produced from peripheral blood mononuclear cells	Heterozygous frameshift mutation in C19orf12 brought by the insertion	[23]
	Episomal plasmids	iPSCs produced from mononuclear cells	No serious adverse events related to CYP-001	[24]
	Episomal plasmids	iPSCs produced from mouse embryonic fibroblast with small molecules	Tenfold increase in reprogramming efficiency	[25]
	Episomal plasmids	iPSCs produced from a peri-infarct area	Endogenous brain repair, reduced inflammation and glial scar formation	[26]
	Episomal plasmids	iPSCs produced from an amyotrophic lateral sclerosis patient’s cell	5-hydroxymethyl cytosine levels increase the reprogramming	[27]
	Circular DNA plasmids	iPSCs produced from B16F10 cells	Did not form teratomas, suppression of tumorigenic abilities	[28]
	mRNA	iPSCs produced from neurons	Purified and differentiated into hair cell-like cells and neurons	[29]
	mRNA	iPSCs produced from urine-derived cells	Generating feeder-free bulk hiPSC lines without genomic abnormalities	[30]
	Small molecules	iPSCs produced from mouse embryonic fibroblasts	Facilitates both in vitro and in vivo alterations in cell fate	[31]
	Small molecules	iPSCs produced from neural stem cells	Melatonin promoted N-iPSC proliferation	[32]
	CRISPR-Cas9	iPSCs produced from skin biopsies	Generate gene-edited hiPSCs from carrying a point mutation	[33]
	Epigenetic modifications	iPSCs produced from mouse fibroblasts	Reconfigurations rapidly propel deterministic reprogramming toward naive pluripotency	[34]
	C9ORF72-mutated	iPSCs produced from fibroblasts and peripheral blood cells	iPSCs and motor neurons derived from the two tissues showed identical properties and features	[35]
	CtIP protein	iPSCs produced from mouse embryonic fibroblast	DNA repair fidelity to both human and mouse iPSCs	[36]
	hiPSC3F-FIB or hiPSC4F-FIB	iPSCs produced from human fibroblasts and fetal neural stem cells	Does not alter subsequent differentiation into neural lineages	[37]
	Integrated at the AAVS1 locus	iPSCs produced from neuron cells with neurogenin 2 transgene	In LOPAC, tau-lowering compounds has been identified	[38]
	OSKM factors, absence of LIF	iPSCs produced from mouse embryonic fibroblasts	No tumor formation but formation of clear hyaline, hypertrophic cartilage	[39]
	Six different reprogramming methods	iPSCs produced from fibroblasts and reprogramed by Lentivirus, Sendai, MiniCircle, Episomal, mRNA, and microRNA	Best results showed by Sendai-virus-based reprogramming	[40]
iPSC Expansion	Stirred based bioreactors	Expansion of macrophages generated from peripheral blood CD34 + cells-derived iPSCs	Highly pure CD45 + CD11b + CD14 + CD163 + cells, act like professional phagocytes	[41]
	Stirred based bioreactors	1 ~ 4 × 107 iPSCs-derived macrophages can be harvested weekly	The ongoing, precise creation of iPSC-Mac populations	[42]
	Vertical-wheel bioreactors	Expansion of human iPSCs as aggregates in single-use bioreactors	Expand iPSCs to expand cells up to 2.3 × 106 (Maximum cell density)	[43]
	Vertical-wheel bioreactors	With a cumulative cell expansion of 1.06 × tenfold in 28 days, the expansion is 30 times in 6 days	Rapid generation of high-quality hiPSCs	[44]
	Vertical-wheel bioreactors with GelMA microcarriers	8-day cell growth that increased 16-fold, differentiation, and immune modulation capacity	Robust, scalable, and cost-effective with translational potential	[45]
	Spinner flask bioreactors	Primary macrophages with cytokine release, phagocytosis, and chemotaxis	Synthesis of genetically altered, iPSC-derived macrophages on a large scale	[46]
	Hydrogel-based 3D culture	Promotes endothelial-network formation and identifies angiogenesis inhibitors	Superior sensitivity and reproducibility over Matrigel	[47]
	Hydrogel-based 3D culture	Fibroblasts formed tiny clusters, spheroids, short segments and on day 20, lengthy segments	The production of closed, inexpensive devices and iPSCs is more rapid, reliable, and scalable	[48]
	Transwell-based 3D culture	In vivo, ex vivo, and in vitro nephrogenic potential, able to produce metabolites that resemble urine	A platform for renal disorders, drug discovery, and human nephrogenesis	[49]
	Multi-culture flasks	Glycogen synthase kinase-3b suppression, CHIR99021 causes a massive proliferation of hiPSC-CMs in vitro (100- to 250-fold)	Expanding hiPSCs for use in tissue engineering and drug screening in a large-scale	[50]
	Chemically defined culture medium	Human skin fibroblasts or peripheral blood mononuclear cells are used to create iPSCs	Differentiation into three embryonic germ layers	[51]
	Chemically defined culture medium	hiPSCs with increased metabolic activity derived from blastocysts or somatic cells	GMP-friendly methods for the manufacturing and processing of therapeutic hiPSC	[52]
	Plate shaker based liquid handler	Cell seeding, splitting, expansion, differentiation image-based multiparametric screening	NPC's neuronal differentiation in 3D midbrain organoids and 2D culture	[53]
	Culture dishes coated with polymer	Create particles with zwitterionic polymer that resemble hyaline cartilaginous tissue and type II collagenopathy	Mass production of chondrocytes and cartilaginous tissues used for drug screening	[54]
Establishment of iPSC Line	Mutagenized iPSC line	CRISPR/Cas9-dependent reprogramming iPSCs	Development of loss-of-function disease models	[55]
	Heterozygous COL1A1 mutation iPSC lines	Karyotype expressed pluripotency markers	Osteogenesis imperfecta disease mechanisms	[56]
	Homozygous/heterozygous iPSC lines	CRISPR-Cas9 dependent reprogramming	Generation of two isogenic iPSC lines	[57]
	KCNA2 mutation iPSC lines	KCNA2 point mutation for produce induces pluripotent stem cells	Expression of pluripotency markers, differentiation into three germ layers	[58]
	Footprint-free iPSC lines	Whole-genome sequencing-based annotated iPSCs lines	Personal Genome Project Canada for personalized iPSC line	[59]
	cGMP-manufactured hiPSC lines	Can produce retinal cells	A human iPSC line that has been used to create transplantable photoreceptors	[60]
	CD34 + hematopoietic cells iPSC lines	CD34 + hematopoietic stem cells from peripheral blood	The production and characterization of three hiPSC lines compatible with GMP	[61]
Process Automation	Fully automated	Microcolonies throughout a 7-day period, sensitivity of 88%, and 98% detection specificity	label-free sensing and mother colony maintenance	[62]
	Fully automated	Retinal pigment epithelial cells are produced using TECAN Fluent automated cell culture	A commercially available platform called end-to-end workflow	[63]
	Automated reprogramming process	Platform for differentiated cells that uses robotics and human involvement	Population-scale personalized iPSC line	[64]
	Automated reprogramming, isolation, and expansion process	Expression of the TRA-1–60 marker for pluripotent stem cells	Commercialized iPSCs line establishment	[65]
	Automated cell culture process	The cell yields, aggregation rates, and expression were higher in non-centrifugation populations	Successfully transferred to independent laboratories	[66]
	Automated cell culture process	Differentiated into dopaminergic neurons, pancreatic cells, and pancreatic hormones	Differentiated into three germ layers	[15]
	Automated quality assessment process	A k-NN classifier with three potential classes has the best accuracy (62,4%) for classification	Automatic evaluation of iPSC colony image quality	[67]
	Biologically inspired AI-based automated process	More adaptable and capable of resolving a wide variety of optimization issues	A necessary simulation is introduced along with the proper model fitting technique	[68]