# Imaging methods to monitor and quantify cell differentiation

**Authors:** Kevin Cheah, Pingjin Chu, Greta Schmidt, Suzanne Scarlata

PMC · DOI: 10.3389/fcell.2025.1584858 · Frontiers in Cell and Developmental Biology · 2025-05-13

## TL;DR

This paper discusses imaging techniques to track and measure how cells change from stem cells to specialized types like neurons and fibroblasts.

## Contribution

The paper introduces real-time imaging methods to monitor dynamic changes during cell differentiation.

## Key findings

- Imaging tools can identify cell lineage during differentiation.
- The methods track changes in mRNA, miRNA, and protein localization.
- These techniques help understand how cells respond to intrinsic and extrinsic factors.

## Abstract

The transition of a cell from a stem to a differentiated state involves an interrelated and complex series of events. These events include dynamic changes in cellular nucleic acid and protein content that are mediated by both intrinsic and extrinsic factors which ultimately lead to differentiation into specific lineage. Quantifying the parameters associated with differentiation and their changes under different conditions would not only allow for a better understanding of this process but also would enable the development of approaches that control differentiation. Here, we describe processes associated with the differentiation of two types of cultured cells, neurons and fibroblasts, and the tools to follow changes in real time. Specifically, we discuss methods to the identify cell lineage, changes in morphology, shifts in specific mRNA and miRNA levels as well as the changes in protein localization, interactions and assemblies that accompany differentiation.

## Full-text entities

- **Genes:** TAFAZZIN (tafazzin, phospholipid-lysophospholipid transacylase) [NCBI Gene 6901] {aka BTHS, CMD3A, EFE, EFE2, G4.5, LVNCX}, JAK2 (Janus kinase 2) [NCBI Gene 3717] {aka JTK10}, MST1 (macrophage stimulating 1) [NCBI Gene 4485] {aka D3F15S2, DNF15S2, HGFL, MSP, NF15S2}, MTOR (mechanistic target of rapamycin kinase) [NCBI Gene 2475] {aka FRAP, FRAP1, FRAP2, RAFT1, RAPT1, SKS}, ILK (integrin linked kinase) [NCBI Gene 3611] {aka HEL-S-28, ILK-1, ILK-2, P59, p59ILK}, Cd44 (CD44 antigen) [NCBI Gene 12505] {aka HERMES, Ly-24, Pgp-1}, SMAD4 (SMAD family member 4) [NCBI Gene 4089] {aka DPC4, JIP, MADH4, MYHRS}, Plcb1 (phospholipase C beta 1) [NCBI Gene 24654] {aka PLC'1, PLCbeta1, Phosphb, RATPHOSPHB}, PIK3CB (phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit beta) [NCBI Gene 5291] {aka P110BETA, PI3K, PI3KBETA, PIK3C1}, Piezo1 (piezo-type mechanosensitive ion channel component 1) [NCBI Gene 234839] {aka 9630020g22, Fam38a, mKIAA0233}, ACTA2 (actin alpha 2, smooth muscle) [NCBI Gene 59] {aka ACTSA, SMDYS}, Ngf (nerve growth factor) [NCBI Gene 310738] {aka Ngfb, beta-NGF}, MAPK1 (mitogen-activated protein kinase 1) [NCBI Gene 5594] {aka ERK, ERK-2, ERK2, ERT1, MAPK2, NS13}, CTNND1 (catenin delta 1) [NCBI Gene 1500] {aka BCDS2, CAS, CTNND, P120CAS, P120CTN, p120}, PPEF1 (protein phosphatase with EF-hand domain 1) [NCBI Gene 5475] {aka PP7, PPEF, PPP7C, PPP7CA}, PLCB1 (phospholipase C beta 1) [NCBI Gene 23236] {aka DEE12, EIEE12, PI-PLC, PLC-154, PLC-I, PLC-beta-1}, TP73 (tumor protein p73) [NCBI Gene 7161] {aka CILD47, P73}, NEUROD1 (neuronal differentiation 1) [NCBI Gene 4760] {aka BETA2, BHF-1, MODY6, NEUROD, T2D, bHLHa3}, SRF (serum response factor) [NCBI Gene 6722] {aka MCM1}, MOB1A (MOB kinase activator 1A) [NCBI Gene 55233] {aka C2orf6, MABKL1B, MATS1, MOB1, MOBK1B, MOBKL1B}, MYH2 (myosin heavy chain 2) [NCBI Gene 4620] {aka CMYO6, CMYP6, IBM3, MYH2A, MYHSA2, MYHas8}, GOLPH3 (golgi phosphoprotein 3) [NCBI Gene 64083] {aka GOPP1, GPP34, MIDAS, Vps74}, RAP1A (RAP1A, member of RAS oncogene family) [NCBI Gene 5906] {aka C21KG, G-22K, KREV-1, KREV1, RAP1, SMGP21}, Smad2 (SMAD family member 2) [NCBI Gene 17126] {aka 7120426M23Rik, Madh2, Madr2, Smad-2, mMad2}, SMAD1 (SMAD family member 1) [NCBI Gene 4086] {aka BSP-1, BSP1, JV4-1, JV41, MADH1, MADR1}, Piezo2 (piezo-type mechanosensitive ion channel component 2) [NCBI Gene 667742] {aka 5930434P17, 9030411M15Rik, 9430028L06Rik, Fam38b, Fam38b2}, Ddr2 (discoidin domain receptor family, member 2) [NCBI Gene 18214] {aka Ntrkr3, tyro10}, Trpv4 (transient receptor potential cation channel, subfamily V, member 4) [NCBI Gene 63873] {aka 0610033B08Rik, OTRPC4, Trp12, VR-OAC, VRL-2, VROAC}, Ptk2 (PTK2 protein tyrosine kinase 2) [NCBI Gene 14083] {aka FADK 1, FAK, FRNK, Fadk, p125FAK}, FURIN (furin, paired basic amino acid cleaving enzyme) [NCBI Gene 5045] {aka FUR, PACE, PCSK3, SPC1}, EGR1 (early growth response 1) [NCBI Gene 1958] {aka AT225, G0S30, KROX-24, NGFI-A, TIS8, ZIF-268}, Smad3 (SMAD family member 3) [NCBI Gene 17127] {aka Madh3}, NANOG (Nanog homeobox) [NCBI Gene 79923], TGFB1 (transforming growth factor beta 1) [NCBI Gene 7040] {aka CAEND1, CED, DPD1, IBDIMDE, LAP, TGF-beta1}, Ddr1 (discoidin domain receptor family, member 1) [NCBI Gene 12305] {aka 6030432F18, CD167a, Cak, Nep, PTK3A}, CAPG (capping actin protein, gelsolin like) [NCBI Gene 822] {aka AFCP, HEL-S-66, MCP}, VTN (vitronectin) [NCBI Gene 7448] {aka V75, VN, VNT}, ERBB2 (erb-b2 receptor tyrosine kinase 2) [NCBI Gene 2064] {aka CD340, HER-2, HER-2/neu, HER2, MLN 19, MLN-19}, UCK2 (uridine-cytidine kinase 2) [NCBI Gene 7371] {aka TSA903, UK, UMPK}, NGF (nerve growth factor) [NCBI Gene 4803] {aka Beta-NGF, HSAN5, NGFB}, MYH14 (myosin heavy chain 14) [NCBI Gene 79784] {aka DFNA4, DFNA4A, FP17425, MHC16, MYH17, NMHC II-C}, Egr1 (early growth response 1) [NCBI Gene 24330] {aka Krox-24, NGFI-A, Ngf1, Ngfi, zif-268}, LTBP1 (latent transforming growth factor beta binding protein 1) [NCBI Gene 4052] {aka ARCL2E}, ZHX2 (zinc fingers and homeoboxes 2) [NCBI Gene 22882] {aka AFR1, RAF}, LAP (Laryngeal adductor paralysis) [NCBI Gene 7939], AKT1 (AKT serine/threonine kinase 1) [NCBI Gene 207] {aka AKT, PKB, PKB-ALPHA, PRKBA, RAC, RAC-ALPHA}, CCN2 (cellular communication network factor 2) [NCBI Gene 1490] {aka CTGF, HCS24, IBP-8, IGFBP8, KMD, NOV2}, PAX6 (paired box 6) [NCBI Gene 5080] {aka AN, AN1, AN2, ASGD5, D11S812E, FVH1}, Acta2 (actin alpha 2, smooth muscle, aorta) [NCBI Gene 11475] {aka 0610041G09Rik, Actvs, SMAalpha, SMalphaA, a-SMA, alphaSMA}, POU5F1 (POU class 5 homeobox 1) [NCBI Gene 5460] {aka OCT3, OCT4, OCT4Borf1, OTF-3, OTF3, OTF4}, CREB1 (cAMP responsive element binding protein 1) [NCBI Gene 1385] {aka CREB, CREB-1}, NTRK1 (neurotrophic receptor tyrosine kinase 1) [NCBI Gene 4914] {aka MTC, TRK, TRK1, TRKA, Trk-A, p140-TrkA}, FN1 (fibronectin 1) [NCBI Gene 2335] {aka CIG, ED-B, FINC, FN, FNZ, GFND}, RBMS3 (RNA binding motif single stranded interacting protein 3) [NCBI Gene 27303], YAP1 (Yes1 associated transcriptional regulator) [NCBI Gene 10413] {aka COB1, YAP, YAP-1, YAP2, YAP65, YKI}, STK4 (serine/threonine kinase 4) [NCBI Gene 6789] {aka KRS2, MST1, YSK3}, MOB1B (MOB kinase activator 1B) [NCBI Gene 92597] {aka MATS2, MOB4A, MOBKL1A}, MYLK3 (myosin light chain kinase 3) [NCBI Gene 91807] {aka MLCK, MLCK2, caMLCK}, APEX1 (apurinic/apyrimidinic endodeoxyribonuclease 1) [NCBI Gene 328] {aka APE, APE1, APEN, APEX, APX, HAP1}, LATS1 (large tumor suppressor kinase 1) [NCBI Gene 9113] {aka WARTS, wts}
- **Diseases:** neurodegenerative diseases (MESH:D019636), neuroblastoma (MESH:D009447), embryonal carcinoma (MESH:D018236), dermal and renal fibrosis (MESH:D005355), cancer (MESH:D009369), hypertrophic scars (MESH:D017439), phototoxicity (MESH:D017484), tissue injury (MESH:D017695), neurological disorders (MESH:D009461), Parkinson's disease (MESH:D010300), Alzheimer's disease (MESH:D000544), liver, kidney and lung fibrosis (MESH:D008103), neurotoxicity (MESH:D020258), schizophrenia (MESH:D012559), epileptic encephalopathy (MESH:D001927)
- **Chemicals:** carbon (MESH:D002244), gold (MESH:D006046), Mg2+ (-), calcium (MESH:D002118), cytosine (MESH:D003596), cytidine (MESH:D003562), RA (MESH:D014212), norepinephrine (MESH:D009638), rhodamine (MESH:D012235), retinol (MESH:D014801), Ag (MESH:D012834), serotonin (MESH:D012701), polyA (MESH:D011061), adenine (MESH:D000225), dbcAMP (MESH:D003994), phorbol esters (MESH:D010703), polydopamine (MESH:C568283), vinyl ether (MESH:C100195), dopamine (MESH:D004298), MnO2 (MESH:C016552), dinitrogen (MESH:D009584), black phosphorus (MESH:D010758), catecholamines (MESH:D002395), acetylcholine (MESH:D000109), Hyaluronic acid (MESH:D006820)
- **Species:** Homo sapiens (human, species) [taxon 9606], Escherichia phage MS2 (no rank) [taxon 12022], Mus musculus (house mouse, species) [taxon 10090], Rattus norvegicus (brown rat, species) [taxon 10116]
- **Cell lines:** NIH-3T3 — Mus musculus (Mouse), Spontaneously immortalized cell line (CVCL_0594), PC12 — Rattus norvegicus (Rat), Rat adrenal gland pheochromocytoma, Cancer cell line (CVCL_0481), SK-N-SH — Homo sapiens (Human), Neuroblastoma, Cancer cell line (CVCL_0531), NT2 — Mus musculus (Mouse), Malignant neoplasms of the mouse mammary gland, Cancer cell line (CVCL_JA57), MEFs — Mus musculus (Mouse), Finite cell line (CVCL_9115), NTERA2 — Homo sapiens (Human), Embryonal carcinoma, Cancer cell line (CVCL_0034), SH-SY5Y — Homo sapiens (Human), Neuroblastoma, Cancer cell line (CVCL_0019)

## Full text

_Full body text omitted from this summary view._ Fetch the complete paper as Markdown: https://tomesphere.com/paper/PMC12106324/full.md

## Figures

11 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12106324/full.md

## References

156 references — full list in the complete paper: https://tomesphere.com/paper/PMC12106324/full.md

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Source: https://tomesphere.com/paper/PMC12106324