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iPSC-Derived MSCs (iMSCs): Derivation, Identity and Consistency

Por Benn Bluestein-Veyra, M.Sc. en Química Orgánica · Actualizado el 20 de octubre de 2026

iPSC-derived mesenchymal stromal cells (iMSCs), also called iPSC-derived mesenchymal stem cells, are made in culture by differentiating induced pluripotent stem cells, and they meet the marker and differentiation criteria applied to MSCs isolated from tissue.^[1][2] Each lot can start from one banked iPSC line, which makes iMSCs a renewable MSC source suited to lot-to-lot consistency.^[3]

This reference covers how iMSCs are derived and characterized, and what side-by-side studies measured against bone marrow MSCs. It also describes master cell banking, iMSC-derived extracellular vesicles and the assays that measure residual pluripotent cells in a differentiated culture.

Understanding iPSC-derived mesenchymal stromal cells as a renewable cell source

Takahashi and colleagues reprogrammed adult human dermal fibroblasts into induced pluripotent stem cells in 2007 with four transcription factors: OCT3/4, SOX2, KLF4 and c-MYC. The cells resembled embryonic stem cells in morphology, proliferation, surface antigens, gene expression and telomerase activity.^[4]

Human iPSCs grow as compact colonies that resemble embryonic stem cell colonies (Fig. 1). They differentiate in vitro and in mice into cell types of all three germ layers, which makes a single iPSC line a starting point for mesenchymal derivation.^[4]

Chen and colleagues pointed to the rarity of MSCs in adult organs, their heterogeneity and the harvest procedures they require as reasons to derive MSCs from iPSCs.^[5] Wruck and colleagues describe the iMSC concept as a way to reduce heterogeneity tied to tissue origin and donor age.^[6]

Derivation routes guide pluripotent cells toward a mesenchymal fate

Dupuis and Oltra grouped published iMSC protocols into five categories: MSC switch, embryoid body formation, specific differentiation, pathway inhibitor and platelet lysate methods (Fig. 2). They also compiled the positive and negative markers used to characterize the resulting cells.^[3]

The routes build on embryonic stem cell work. In 2005, Barberi and colleagues described culture conditions that gave pure mesenchymal precursors from human embryonic stem cells, which differentiated into fat, cartilage, bone and skeletal muscle cells.^[7]

Embryoid body formation

In embryoid body protocols, iPSCs are first aggregated into three-dimensional bodies, and the adherent cells that grow out are expanded in MSC medium.^[3] Ababneh and colleagues compared an embryoid body protocol using retinoic acid with four monolayer protocols, and every protocol gave cells expressing MSC markers and lacking pluripotency markers.^[8]

Monolayer differentiation

Hynes and colleagues cultured iPSC colonies directly in MSC medium for 2 weeks and then used serial passaging to select fast-growing fibroblastic cells. They obtained MSC-like cells from iPSC lines reprogrammed from gingiva, periodontal ligament and lung.^[1]

Chen and colleagues cultured iPSCs in serum-free medium with SB431542, an inhibitor of the transforming growth factor-β pathway, which gave epithelial-like monolayers. Transfer to conventional MSC medium then drove an epithelial-to-mesenchymal transition into MSC-like cells.^[5]

Liu and colleagues plated single iPSCs on thin fibrillar type I collagen coatings in medium with ascorbate and dexamethasone. Within 10 d, homogeneous colonies of spindle-shaped cells formed that expressed CD73, CD90, CD105, CD146 and CD166.^[9]

Neural crest and mesoderm intermediates

Neural crest cells contribute to the craniofacial skeleton and can give rise to mesenchymal cells.^[10] Lee and colleagues derived neural crest stem cells from human embryonic stem cells that differentiated toward smooth muscle, adipogenic, osteogenic and chondrogenic cells.^[11]

Menendez and colleagues activated canonical Wnt signaling while blocking the Activin A and Nodal pathway, converting pluripotent cells into p75, HNK1 and AP2 positive neural crest-like cells over 12 to 14 d. These cells gave mesenchymal precursors that formed osteocytes, chondrocytes and adipocytes.^[12]

Fukuta and colleagues combined a GSK3β inhibitor and a TGFβ inhibitor in chemically defined medium and induced neural crest cells at 70 to 80% efficiency. Culture in a defined MSC medium then produced cells with MSC-like differentiation characteristics.^[10]

Eto and colleagues derived iMSCs through mesoderm-like and neuroepithelium-like intermediates, and both populations self-renewed and were multipotent.^[13]

Characterizing iMSCs against the ISCT minimal criteria

The ISCT minimal criteria define MSCs by plastic adherence, expression of CD105, CD73 and CD90, absence of CD45, CD34, CD14 or CD11b, CD79α or CD19 and HLA-DR, and differentiation to osteoblasts, adipocytes and chondroblasts in vitro.^[14] iMSC studies test their cells against this same panel.^[3]

Kang and colleagues found two iMSC lines positive for CD29, CD44, CD73, CD90 and CD105 and negative for CD11b, CD14, CD31, CD34, CD45 and HLA-DR, matching bone marrow MSCs.^[2] CD73 is the ecto-5′-nucleotidase encoded by NT5E, a membrane enzyme that converts extracellular nucleotides to nucleosides (Fig. 3).^[15]

Pluripotency markers form a second, negative panel. Hynes and colleagues confirmed that iMSCs lacked TRA-1-60, TRA-1-81 and alkaline phosphatase, and Kang and colleagues found OCT4, SOX2, GDF3 and LIN28A expression absent in iMSCs and their differentiated progeny.^[1][2]

Surface marker sorting

Lian and colleagues found CD105 highly expressed on embryonic stem cell-derived MSCs and CD24 on the parent stem cells. CD105-positive, CD24-negative isolates had typical MSC gene expression and were highly correlated with each other, with r² above 0.90.^[16]

Diederichs and Tuan derived MSC-like progenitors by four methods from an iPSC line reprogrammed from bone marrow stromal cells and compared them with the parental cells. The derived cells met the morphology, marker and trilineage criteria and showed a distinct pattern of mesenchymal and pluripotency gene expression.^[17]

How iMSCs compare with tissue-derived MSCs in growth, lineage and secretome

Side-by-side studies compare iMSCs with bone marrow MSCs and, in some designs, with parental cells from the same donor (Fig. 4).^[2][17] The table summarizes the attributes measured most often, and the sections below give the study behind each finding.

Proliferation and senescence

Kang and colleagues saw faster proliferation in both iMSC lines than in bone marrow MSCs.^[2] Ababneh and colleagues found proliferation comparable to or above that of bone marrow MSCs in four of five protocols, with lower reactive oxygen species in all iMSCs and lower senescence in four.^[8]

Lineage preference

Kang and colleagues reported adequate osteogenic and chondrogenic capacity in iMSCs and less adipogenic potential.^[2] Chen and colleagues likewise saw robust osteogenesis and chondrogenesis with limited adipogenesis, a pattern also reported for fetal MSCs and embryonic stem cell-derived MSCs.^[5]

Ababneh and colleagues found enhanced RUNX2 and reduced adipsin in embryoid body-derived iMSCs, so the protocol shapes lineage balance.^[8] Hynes and colleagues saw differentiation capacity vary between iPSC lines, and only cells from the periodontal ligament line formed mineralized structures after implantation in mice.^[1]

Secretome and co-culture assays

Spitzhorn and colleagues measured an iMSC secretome that included angiogenin, DKK-1, IL-8, PDGF-AA, osteopontin, SERPINE1 and VEGF. The profile was similar to that of fetal and adult MSCs, which points to paracrine signaling as a shared feature.^[18]

T cell co-culture results vary between studies. Frobel and colleagues found iMSCs less able than primary MSCs to suppress T cell proliferation, while Ng and colleagues found suppression of mitogen-stimulated mononuclear cells similar to the parental fibroblasts, with raised IDO1 and IL6 transcripts.^[19][20]

Buitrago and colleagues reported that iMSCs regulated lymphocyte proliferation and shifted monocytes toward an anti-inflammatory phenotype to a degree comparable with umbilical cord MSCs. They also measured lower TNFα and higher IL-10 in these co-cultures.^[21]

Reprogramming resets the epigenetic age of iPSC-derived MSCs

Long-term culture of MSCs brings reproducible senescence-associated DNA methylation changes, enriched in nonpromoter regions of developmental genes. Koch and colleagues showed that reprogramming into iPSCs prevented almost the entire set of these changes.^[22]

Lapasset and colleagues reprogrammed senescent and centenarian fibroblasts into iPSCs with reset telomere size, gene expression, oxidative stress and mitochondrial metabolism. Cells redifferentiated from these iPSCs were described as fully rejuvenated.^[23]

Frobel and colleagues reprogrammed MSCs into iPSCs and differentiated them back to MSCs. Tissue-specific, senescence-associated and age-related methylation patterns were erased, and the iMSCs reacquired senescence-associated methylation during expansion while staying rejuvenated in age-related methylation.^[19]

Spitzhorn and colleagues made iMSCs from iPSCs of fetal and aged adult MSCs. Irrespective of donor age, the iMSCs acquired a rejuvenation signature including INHBE, DNMT3B, CDKN1C and GCNT2, and shared more genes with fetal MSCs than with adult MSCs.^[18]

Frobel and colleagues also found that iPSC methylation profiles kept donor-specific features.^[19] In a meta-analysis of MSC transcriptomes, Wruck and colleagues linked heterogeneity mainly to tissue origin and proposed iMSCs as a way to bypass tissue- and age-related variation.^[6]

A master iPSC bank supports consistency and scale

Dupuis and Oltra describe iPSCs as a reliable, unlimited source for generating MSC lines.^[3] Sullivan and colleagues set out consensus recommendations on the minimum dataset of critical quality attributes and assays for a clinical-grade iPSC line, so lines from different donors and facilities can be compared.^[24]

Umekage and colleagues describe the iPSC stock at the Center for iPS Cell Research and Application in Kyoto, made from peripheral blood or cord blood of consenting HLA-homozygous donors. In 2019 the stock offered 21 clones from 5 donors.^[25]

Lian and colleagues generated karyotypically stable, identical batches of embryonic stem cell-derived MSCs without virus, mouse cells or serum.^[16] Diederichs and Tuan noted that the proliferative capacity of pluripotent cells opens the possibility of large, uniform MSC batches.^[17]

Rogers and colleagues expanded iMSCs on degradable gelatin methacryloyl microcarriers in vertical wheel bioreactors, reaching 8.8 to 16.3-fold expansion over 8 d. Microcarrier digestion released cells at over 95% viability with immunomodulatory potential in vitro equal to or above monolayer-expanded iMSCs.^[26]

iMSC-derived extracellular vesicles are an active research direction

Kim and colleagues established iPSCs from Wharton’s jelly MSCs, differentiated them into iMSCs, and collected exosomes from the culture supernatant of both cell types. Both preparations showed the basic characteristics of exosomes and were taken up by keratinocytes and dermal fibroblasts in culture, and the authors highlighted iMSCs as producer cells for exosomes.^[27]

Buitrago and colleagues reported that iMSC vesicles inhibited T cell proliferation and influenced macrophage polarization in vitro, similar to their parent cells.^[21]

Residual pluripotent cell testing is a defined quality attribute

Tano and colleagues describe the fraction of undifferentiated cells as a quality attribute of any cell preparation derived from pluripotent stem cells. Assays for it are designed to find a few iPSCs among a large number of differentiated cells.^[28]

Their culture assay on laminin-521 in Essential 8 medium amplifies trace iPSCs into countable colonies. It detected iPSCs spiked into primary MSCs at 0.001 to 0.01%, and it detected undifferentiated cells during iMSC differentiation from iPSCs.^[28]

Kuroda and colleagues built a droplet digital PCR assay for LIN28 transcripts that detected 0.001% iPSCs in cardiomyocytes, one cell in 10⁵. LIN28 expression was very low in the adult human tissues they tested.^[29] LIN28A encodes an RNA-binding regulator of self-renewal in embryonic stem cells.^[30]

Yasuda and colleagues ran an international multisite evaluation of a one-step RT-ddPCR assay for ESRG, LINC00678 and LIN28A, with iPSCs spiked into iPSC-derived cardiomyocytes. The assay was sensitive and robust, and most variation between laboratories came from the spiking step.^[31]

OCT4, encoded by POU5F1, is a POU homeodomain transcription factor with a key role in embryonic development and stem cell pluripotency (Fig. 5).^[32] Kang and colleagues reported OCT4 transcripts absent from iMSCs, which makes OCT4 one of the negative markers in iMSC characterization.^[2]

Where to go from here

The ISCT minimal criteria reference explains the marker panel every iMSC study measures against, and the MSC source tissues reference compares bone marrow, adipose, cord and placental cells. The stem cell types reference places iPSCs among the other stem cell classes, and the single-cell heterogeneity reference shows how varied a tissue MSC culture can be.

The MSC secretome reference covers conditioned medium and vesicle fractions, and the exosome biochemistry and surface receptor chemistry references continue from cells to vesicles. Verified practices can request lot documentation, and our team is glad to discuss the methods described on this page.