# Progression in the In Vitro Macrophage Expansion

**Authors:** Yunpeng Wei, Jingzhao Yang, Wenhong Zu, Mengran Wang, Yong Zhao

PMC · DOI: 10.1155/jimr/9994439 · 2025-04-28

## TL;DR

This review discusses how macrophages can be expanded in the lab, highlighting various methods and the need for further research to improve efficiency.

## Contribution

The paper provides a comprehensive summary of factors and mechanisms that promote macrophage proliferation in vitro.

## Key findings

- Macrophage proliferation in vitro can be stimulated by cytokines, small molecules, and other factors.
- Current methods for macrophage expansion are limited by heterogeneity and poor understanding of regulatory mechanisms.
- Further research is needed to develop efficient strategies for large-scale macrophage expansion.

## Abstract

Macrophages play essential roles in homeostasis and disease, and they were considered terminally differentiated cells that cannot proliferate. However, growing evidence shows that macrophages can self-renew in homeostasis and multiple pathological states in vivo and artificial induction in vitro. With the rise of immune cell therapy based on macrophages, large-scale in vitro expansion of macrophages has become more and more urgent. However, the proliferation of macrophages in vitro is still inefficient because of the heterogeneity of macrophages, complicated crosstalk between macrophages and their microenvironments, and poor understanding of macrophage proliferation regulations. In this review, we summarized the discoveries known to stimulate macrophage proliferation in vitro, including cytokines, small molecule compounds, metabolites, the composition of pathogens and apoptotic cells, natural product extracts, gene editing, and other factors, as well as related mechanisms. It can be concluded that the promotion of macrophage proliferation in vitro covers various approaches and mechanisms. However, it is still necessary to test more strategies and learn more macrophage proliferation mechanisms to achieve large-scale engineering expansion of macrophages in vitro.

## Full-text entities

- **Genes:** Crtc2 (CREB regulated transcription coactivator 2) [NCBI Gene 74343] {aka 4632407F12Rik, Torc2}, Csf1r (colony stimulating factor 1 receptor) [NCBI Gene 12978] {aka CD115, CSF-1R, Csfmr, Fim-2, Fim2, Fms}, Rictor (RPTOR independent companion of MTOR, complex 2) [NCBI Gene 78757] {aka 4921505C17Rik, 6030405M08Rik, AVO3, D530039E11Rik}, S1pr1 (sphingosine-1-phosphate receptor 1) [NCBI Gene 13609] {aka Edg1, Lpb1, S1p, S1p1}, Nfkb1 (nuclear factor of kappa light polypeptide gene enhancer in B cells 1, p105) [NCBI Gene 18033] {aka NF-KB1, NF-kappaB, NF-kappaB1, p105, p50, p50/p105}, Mapk14 (mitogen-activated protein kinase 14) [NCBI Gene 26416] {aka CSBP2, Crk1, Csbp1, Mxi2, PRKM14, PRKM15}, Npff (neuropeptide FF-amide peptide precursor) [NCBI Gene 54615], Nr1h3 (nuclear receptor subfamily 1, group H, member 3) [NCBI Gene 22259] {aka LXR, RLD1, Unr1}, Csf1 (colony stimulating factor 1 (macrophage)) [NCBI Gene 12977] {aka BAP025, Csfm, MCSF, Mhdabap25, PG-M-CSF, op}, Spp1 (secreted phosphoprotein 1) [NCBI Gene 20750] {aka 2AR, Apl-1, BNSP, BSPI, Bsp, ETA-1}, Bcl2 (B cell leukemia/lymphoma 2) [NCBI Gene 12043] {aka Bcl-2, C430015F12Rik, D630044D05Rik, D830018M01Rik}, Gsk3b (glycogen synthase kinase 3 beta) [NCBI Gene 56637] {aka 7330414F15Rik, 8430431H08Rik, GSK-3, GSK-3beta, GSK3}, Rheb (Ras homolog enriched in brain) [NCBI Gene 19744] {aka Rheb1}, Hras (Hras proto-oncogene, GTPase) [NCBI Gene 15461] {aka H-ras, Ha-ras, Harvey-ras, Hras-1, Hras1, Kras2}, Cxcl2 (C-X-C motif chemokine ligand 2) [NCBI Gene 20310] {aka CINC-2a, GROb, Gro2, MIP-2, MIP-2a, Mgsa-b}, Sirt1 (sirtuin 1) [NCBI Gene 93759] {aka SIR2L1, Sir2, Sir2a, Sir2alpha}, Csf2 (colony stimulating factor 2) [NCBI Gene 116630] {aka Gm-csf, Gmcsf}, Cdh13 (cadherin 13) [NCBI Gene 12554] {aka 4932416G01Rik, Cdht, Tcad}, Tnf (tumor necrosis factor) [NCBI Gene 21926] {aka DIF, TNF-a, TNF-alpha, TNFSF2, TNFalpha, Tnfa}, Msr1 (macrophage scavenger receptor 1) [NCBI Gene 20288] {aka MRS-A, MSR, MSR-A, SR-A, SR-AI, SR-AII}, Tgfb1 (transforming growth factor, beta 1) [NCBI Gene 21803] {aka TGF-beta1, TGFbeta1, Tgfb, Tgfb-1}, Klf4 (Kruppel-like transcription factor 4 (gut)) [NCBI Gene 16600] {aka EZF, Gklf, Zie}, Calm2 (calmodulin 2) [NCBI Gene 12314] {aka 1500001E21Rik, Cam2, CamC}, Dnase2a (deoxyribonuclease II alpha) [NCBI Gene 13423] {aka Dnase2}, Hnrnpa1 (heterogeneous nuclear ribonucleoprotein A1) [NCBI Gene 15382] {aka HDP-1, Hdp, Hnrpa1, hnRNP A1, hnrnp-A1}, Pik3r1 (phosphoinositide-3-kinase regulatory subunit 1) [NCBI Gene 18708] {aka PI3K, p50alpha, p55alpha, p85alpha}, Tlr9 (toll-like receptor 9) [NCBI Gene 81897], Mir124a-3 (microRNA 124a-3) [NCBI Gene 723951] {aka Mirn124a-3, mir-124-3, mir-124a-3}, Sra1 (steroid receptor RNA activator 1) [NCBI Gene 24068] {aka Sra, Srap, Straa1, Strra1}, Myc (Myc proto-oncogene, bHLH transcription factor) [NCBI Gene 17869] {aka Myc2, Niard, Nird, bHLHe39}, Prkdc (protein kinase, DNA activated, catalytic polypeptide) [NCBI Gene 19090] {aka DNA-PKcs, DNAPDcs, DNAPK, DNPK1, DOXNPH, HYRC1}, Bhlhe41 (basic helix-loop-helix family, member e41) [NCBI Gene 79362] {aka 6430520M22Rik, Bhlhb2l, Bhlhb3, DEC2, Sharp1}, Igf2 (insulin-like growth factor 2) [NCBI Gene 16002] {aka Igf-2, Igf-II, M6pr, Mpr, Peg2}, Igf1 (insulin-like growth factor 1) [NCBI Gene 16000] {aka C730016P09Rik, Igf-1, Igf-I}, Tgfb2 (transforming growth factor, beta 2) [NCBI Gene 21808] {aka Tgf-beta2, Tgfb-2}, Pparg (peroxisome proliferator activated receptor gamma) [NCBI Gene 19016] {aka Nr1c3, PPAR-gamma, PPAR-gamma2, PPARgamma, PPARgamma2}, Map3k3 (mitogen-activated protein kinase kinase kinase 3) [NCBI Gene 26406] {aka MAPKKK3, Mekk3, mKIAA4031}, Cd14 (CD14 antigen) [NCBI Gene 12475], Hsh2d (hematopoietic SH2 domain containing) [NCBI Gene 209488] {aka ALX, Hsh2}, Smad3 (SMAD family member 3) [NCBI Gene 17127] {aka Madh3}, Cdkn1b (cyclin dependent kinase inhibitor 1B) [NCBI Gene 12576] {aka Kip1, p27, p27Kip1}, Cdkn1a (cyclin dependent kinase inhibitor 1A) [NCBI Gene 12575] {aka CAP20, CDKI, CIP1, Cdkn1, P21, SDI1}, Mafb (MAF bZIP transcription factor B) [NCBI Gene 16658] {aka Kreisler, Krml, Krml1, kr}, Ccnd1 (cyclin D1) [NCBI Gene 12443] {aka CycD1, Cyl-1, PRAD1, bcl-1, cD1}, Fos (Fos proto-oncogene, AP-1 transcription factor subunit) [NCBI Gene 14281] {aka D12Rfj1, c-fos, cFos}, Maf (MAF bZIP transcription factor) [NCBI Gene 17132] {aka 2810401A20Rik, A230108G15Rik, c-maf}, Gpr132 (G protein-coupled receptor 132) [NCBI Gene 56696] {aka G2a}, Spon1 (spondin 1, (f-spondin) extracellular matrix protein) [NCBI Gene 233744] {aka D330035F22Rik, FSP}, Igf1 (insulin-like growth factor 1) [NCBI Gene 24482] {aka IGF}, Csf2 (colony stimulating factor 2 (granulocyte-macrophage)) [NCBI Gene 12981] {aka CSF, Csfgm, GMCSF, Gm-CSf, MGI-IGM}, Il1rn (interleukin 1 receptor antagonist) [NCBI Gene 16181] {aka F630041P17Rik, IL-1ra}, Mmp12 (matrix metallopeptidase 12) [NCBI Gene 17381] {aka MME, Mmel}, Hif1a (hypoxia inducible factor 1, alpha subunit) [NCBI Gene 15251] {aka HIF-1-alpha, HIF1-alpha, HIF1alpha, MOP1, bHLHe78}, Adipoq (adiponectin, C1Q and collagen domain containing) [NCBI Gene 11450] {aka 30kDa, APN, Acdc, Acrp30, Ad, Adid}, Bhlhe40 (basic helix-loop-helix family, member e40) [NCBI Gene 20893] {aka Bhlhb2, C130042M06Rik, CR8, Clast5, Dec1, Sharp2}, Akt1 (Akt serine/threonine kinase 1) [NCBI Gene 11651] {aka Akt, LTR-akt, PKB, PKB/Akt, PKBalpha, Rac}, Mapk1 (mitogen-activated protein kinase 1) [NCBI Gene 26413] {aka 9030612K14Rik, ERK, Erk2, MAPK2, PRKM2, Prkm1}, Ctnnb1 (catenin beta 1) [NCBI Gene 12387] {aka Bfc, Catnb, Mesc}, Mapk8 (mitogen-activated protein kinase 8) [NCBI Gene 26419] {aka JNK, JNK1, Prkm8, SAPK1}, Nr0b2 (nuclear receptor subfamily 0, group B, member 2) [NCBI Gene 23957] {aka SHP, SHP-1, Shp1}
- **Diseases:** tumorigenesis (MESH:D063646), nonsmall cell lung cancer (MESH:D002289), solid (MESH:D018250), AMs (MESH:D055501), toxicity (MESH:D064420), diabetes (MESH:D003920), glioblastoma (MESH:D005909), tumorigenic (MESH:D002471), arthritis (MESH:D001168), atherosclerosis (MESH:D050197), pain (MESH:D010146), acute respiratory distress syndrome (MESH:D012128), inflammation (MESH:D007249), colorectal cancer (MESH:D015179), hepatic metastasis (MESH:D009362), tumor (MESH:D009369), hepatocellular carcinoma (MESH:D006528), infections (MESH:D007239), Hyperoxia (MESH:D018496)
- **Chemicals:** LTD4 (MESH:D017998), heavy metal (MESH:D019216), ROS (MESH:D000077154), Eicosapentaenoic Acid (MESH:D015118), thymidine (MESH:D013936), sphingolipid (MESH:D013107), dextran (MESH:D003911), lignin (MESH:D008031), disaccharide (MESH:D004187), Arachidonic Acid (MESH:D016718), hyaluronan (MESH:D006820), CpG-ODN 1668 (-), carbon dioxide (MESH:D002245), Glycol (MESH:D006018), Dexamethasone (MESH:D003907), ATP (MESH:D000255), LTB5 (MESH:C040212), lactate (MESH:D019344), lipid (MESH:D008055), LPS (MESH:D008070), oxygen (MESH:D010100), Urea (MESH:D014508), fatty acid (MESH:D005227), 3H (MESH:D014316), NAD+ (MESH:D009243), tricarboxylic acid (MESH:D014233), C1P (MESH:C065576), RvD1 (MESH:C518399), BPA (MESH:C006780), Heparan Sulfate (MESH:D006497), nucleotides (MESH:D009711), calcium (MESH:D002118), sugars (MESH:D000073893), glycosaminoglycan (MESH:D006025), Metformin (MESH:D008687), glycolaldehyde (MESH:C010972), NO (MESH:D009569), PBS (MESH:D007854), lyso-PC (MESH:D008244), Heparin (MESH:D006493), PGE2 (MESH:D015232), LTB4 (MESH:D007975), CpG oligodeoxynucleotides (MESH:C408982), Erythromycin (MESH:D004917), Cadmium (MESH:D002104), AGEs (MESH:D017127), steroids (MESH:D013256), D-glucuronic acid (MESH:D020723), N-acetylglucosamine (MESH:D000117), morphine (MESH:D009020), PGE3 (MESH:C033246), Polysaccharides (MESH:D011134)
- **Species:** Homo sapiens (human, species) [taxon 9606], Mycoplasmopsis pulmonis (species) [taxon 2107], Oryctolagus cuniculus (domestic rabbit, species) [taxon 9986], Rattus norvegicus (brown rat, species) [taxon 10116], Mus musculus (house mouse, species) [taxon 10090], Metamycoplasma arthritidis (species) [taxon 2111], Lactarius deliciosus (species) [taxon 55514]
- **Cell lines:** J774.A1 — Mus musculus (Mouse), Mouse reticulum cell sarcoma, Cancer cell line (CVCL_0358), U937 — Homo sapiens (Human), Adult acute monocytic leukemia, Cancer cell line (CVCL_0007), THP1 — Homo sapiens (Human), Childhood acute monocytic leukemia, Cancer cell line (CVCL_0006), /c — Mus musculus (Mouse), Hepatocellular carcinoma of the mouse, Cancer cell line (CVCL_9103), Jurkat — Homo sapiens (Human), Childhood T acute lymphoblastic leukemia, Cancer cell line (CVCL_0065), NIH 3T3 — Mus musculus (Mouse), Spontaneously immortalized cell line (CVCL_0594), L929 — Mus musculus (Mouse), Spontaneously immortalized cell line (CVCL_AR58), RAW 264.7 — Mus musculus (Mouse), Mouse leukemia, Cancer cell line (CVCL_0493), C3H/HeJ — Mus musculus (Mouse), Embryonic stem cell (CVCL_2H83)

## Figures

1 figure with captions in the complete paper: https://tomesphere.com/paper/PMC12052461/full.md

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Source: https://tomesphere.com/paper/PMC12052461