Human Plasma-Like Medium | Thermo Fisher Scientific

The HPLM solution

Gibco HPLM contains the same salt concentrations found in human plasma, as well as the same concentrations of over 60 polar metabolites, such as amino acids, nucleic acids, sugars, and small organic acids. In resembling the natural cellular environment found in the body, HPLM helps provide researchers the ability to study the impact of physiologically relevant cell media on their specific applications.

HPLM supplemented with fetal bovine serum (FBS) can support cell growth and viability comparable to those of conventional FBS-supplemented basal media formulations. For most cell lines, adaptation is not required to transition from conventional medium to HPLM.

HPLM is beneficial to your cell culture experiments in several ways:

Physiologically relevant—formulated with more than 60 polar metabolites and salt concentrations that resemble the natural cellular environment found in the body
Peer reviewed—extensive research publications completed using HPLM formulation
Easy to use—direct replacement for your current media when supplemented with FBS

Meet the inventor of HPLM

Watch Jason's interview

The inventor of human plasma-like medium is Jason R. Cantor.

As a postdoc at the Whitehead Institute/MIT in Cambridge, Jason set out to create what would become human plasma-like medium (HPLM), a physiologic medium designed to more closely reflect the metabolic composition of human blood, thus permitting the study of cultured cells in biochemical conditions with greater relevance to human physiology.

Cantor reported his development and initial studies using HPLM in early 2017 (Cell). Read his publication here: Physiologic Medium Rewires Cellular Metabolism and Reveals Uric Acid as an Endogenous Inhibitor of UMP Synthase.

We are proud to work with Jason to bring this innovation to market, and excited by the immense possibilities that HPLM could bring across diverse areas of the scientific community. As Jason notes, "The recent development of physiologic media, like other efforts designed to address the modeling capacity of cell culture, holds immense potential to improve understanding and interpretation of diverse biological and pharmacological studies." Read more from his 2019 commentary here: The Rise of Physiologic Media .

Jason is listed as an inventor on a patent application for HPLM assigned to Whitehead Institute.

Research results using HPLM

Performance
Functionality
Customer stories
Webinar
White paper

Research has shown that cellular performance is impacted by the use of HPLM, indicating that physiologic media can help increase the relevance of results from physiological studies.

HPLM rewires cellular metabolism

Graphical abstract summary: “Among the most prominent was an inhibition of de novo pyrimidine synthesis—an effect traced to uric acid, which is 10-fold higher in the blood of humans than of mice and other non-primates. We find that uric acid directly inhibits uridine monophosphate synthase (UMPS) and consequently reduces the sensitivity of cancer cells to the chemotherapeutic agent 5-fluorouracil. Thus, media that better recapitulates the composition of human plasma reveals unforeseen metabolic wiring and regulation, suggesting that HPLM should be of broad utility.”

Reproduced with permission from: Cantor JR, Abu-Remaileh M, Kanarkek N et al. (2017) Physiologic medium rewires cellular metabolism and reveals uric acid as an endogenous inhibitor of UMP synthase. Cell 169: 258–272.E17. doi: 10.1016/j.cell.2017.03.023

HPLM improves T lymphocyte activation

Graphical abstract summary: “Among the most prominent was an inhibition of de novo pyrimidine synthesis—an effect traced to uric acid, which is 10-fold higher in the blood of humans than of mice and other non-primates. We find that uric acid directly inhibits uridine monophosphate synthase (UMPS) and consequently reduces the sensitivity of cancer cells to the chemotherapeutic agent 5-fluorouracil. Thus, media that better recapitulates the composition of human plasma reveals unforeseen metabolic wiring and regulation, suggesting that HPLM should be of broad utility.”

Reproduced with permission from: Cantor JR, Abu-Remaileh M, Kanarkek N et al. (2017) Physiologic medium rewires cellular metabolism and reveals uric acid as an endogenous inhibitor of UMP synthase. Cell 169: 258–272.E17. doi: 10.1016/j.cell.2017.03.023

The functional characteristics of cells, including morphology and growth, in HPLM are comparable to those in conventional basal media formulations.

Human plasma-like medium maintains cell morphology

2-panel brightfield microscopic views showing that MCF7 cells growing in either DMEM or HPLM exhibit the same morphologies

Figure 1. Gibco HPLM supports MCF7 cell culture. MCF7 breast adenocarcinoma cells were cultured in DMEM (Cat. No. 10566016) or Human Plasma-Like Medium (HPLM, Cat. No. A4899101) supplemented with 10% FBS (Cat. No. A3840101).

Figure 2. Gibco HPLM supports HeLa cell culture. HeLa human cervical adenocarcinoma cells were cultured in DMEM (Cat. No. 11965092) or Human Plasma-Like Medium (HPLM, Cat. No. A4899101) supplemented with 10% FBS (Cat. No. A3840101).

2-panel brightfield microscopic views showing that LNCaP cells growing in either DMEM or HPLM exhibit the same morphologies

Figure 3. Gibco HPLM supports LNCaP cell culture. LNCaP human metastatic prostate carcinoma cells were cultured in RPMI 1640 (left; Cat. No. 61870036) or HPLM (right; Cat. No. A4899101) supplemented with 10% FBS (Cat. No. A3840101).

Human plasma-like medium maintains cell growth

4-panel bar charts showing comparable doubling times for 4 cell lines in DMEM and in HPLM

Figure 4. Gibco HPLM supports comparable growth rates in continuous culture. Tumor cell lines MCF7, HeLa, A549, and THP-1 were grown in DMEM (blue) or HPLM (red) supplemented with 10% FBS (Cat. No. A3840101) for five passages. Cell number was assessed at the end of each passage and used to calculate the average population doubling time for each culture.

Human plasma-like medium supports cell growth in multiple cell types

Table 1. Cell types successfully grown in human plasma-like medium

Cell type tested	Cell origin
A375, adherent	Malignant melanoma
A549, adherent	Lung carcinoma
HCT116, adherent	Colorectal carcinoma
HeLa, adherent	Cervical adenocarcinoma
Jurkat, suspension	T cell leukemia
LNCaP, adherent	Metastatic prostate carcinoma
MCF-7, adherent	Metastatic breast cancer
MDA-MB-231, adherent	Metastatic breast cancer
NK, primary, suspension	Lymphocyte, blood
NOMO-1, suspension	Monoblastic/monocytic leukemia
PLB-985, suspension	Myeloid leukemia
Sp2, suspension	Mouse B lymphocyte
THP-1, suspension	Monocytic leukemia
U-2 OS, adherent	Osteosarcoma
WM115, adherent	Malignant melanoma

The following customer stories feature the work of our Gibco HPLM product testers

“We used the HPLM media with cancer cell lines from different tissues and different mutation background. Our overall impression of HPLM is quite positive. HPLM media was suitable for all the tested cancer cell lines. Cells were easily and quickly adapted to the HPLM without affecting viability and cell culture performance. However, we detected relevant impact of cell proliferation, metabolism and mitochondrial function in cells grown in HPLM as compared with classical media. After our experience using HPLM in cancer cell models we consider it as the best choice to get more physiological data.”

Omar Torres-Quesada

PhD, Postdoctoral Researcher at University of Innsbruck

Austria

“Immunometabolism is an exciting area of scientific investigation that has immense potential for the development of new therapeutics. It is becoming clear that how immune cells rewire their metabolism after activation depends on what nutrients are available and in what quantities. Since we study metabolite transport in primary human immune cells, we find HPLM media to be essential for modeling metabolic flux in the most physiologically relevant way possible. We have extensively vetted the use of HPLM across CD4 T helper cell lineages and find that it supports both proliferation and effector function."

Justin Rettenmaier,

Associate Director at Jnana Therapeutics

Boston, MA

"Standard cell culture media, including the Dulbecco’s Modified Eagle Medium (DMEM), contain a non-physiological excess of nutrients, including glucose and glutamine. At the same time, they are low on uric acid. Nutrient availability, however, is a most critical factor for the regulation of mTOR calling for a careful consideration of culture conditions. Therefore, we employed Human Plasma Like Medium (HPLM) to gain a better understanding of mTORC1 regulation under physiological nutrient abundance. In line with mTOR’s intricate connection to nutrient sensing pathways, mTORC1 activity, as measured by Thr389 phosphorylation of S6K, was lower when U2OS cells were cultured in HPLM instead of DMEM. In HPLM media and irrespective of any treatment, AKT and the glucose-sensing AMPK were activated. Most importantly, however, we found that p53 was required to sustain low mTORC1 activity under HPLM culture conditions irrespective of Nutlin-3a treatment. Together, our investigation using HPLM revealed a nutrient-dependent role of p53 in mTORC1 inhibition."

Martin Fischer

Principal Investigator at Leibniz Institute on Aging

Germany

Webinar: Physiologic medium to study human cell biology

Environmental factors influence human cell physiology and can also affect drug efficacy, but existing model systems used to study human cells have limitations for understanding these contributions. In this webinar, Dr. Jason Cantor will discuss the initial development and use of human plasma-like medium (HPLM), a physiologic medium designed to more closely reflect the metabolic composition of human blood. By examining human cancer cell lines in HPLM versus traditional media, Dr. Cantor and colleagues have recently shown that HPLM has widespread effects on metabolism and gene essentiality, and further, that HPLM can be used to reveal new insights into metabolic regulation and drug efficacy.

Speaker: Jason R. Cantor, Investigator, Morgridge Institute for Research, Assistant Professor of Biochemistry, University of Wisconsin-Madison

Webinar highlights:

Complete media, the workhorses of cell culture studies, typically consist of a basal medium that poorly resembles conditions encountered in the human body and a largely undefined serum supplement
Human Plasma-Like Medium (HPLM) is a relatively new physiologic basal medium designed to more closely reflect the metabolic composition of human blood
The development of physiologic media, like other efforts designed to address the modeling capacity of cell culture, holds immense potential to improve understanding and interpretation of biological and pharmacological studies

Watch webinar

Assessment of cancer spheroid formation in Human Plasma-Like Medium

Human Plasma-Like Medium (HPLM) is a cell growth formulation designed to mimic the metabolic profile of human plasma to help maintain the physiological state of cells.

We demonstrate the capability of HPLM to support the growth of cancer spheroids in several well-studied cancer cell lines

Microscopy images of spheroids for different cell types grown in standard RPMI or HPLM medium

Figure 5. HPLM supports 3D spheroid formation. Representative images of spheroids from various cell types cultured in RPMI standard medium and Human Plasma-Like Medium (HPLM) that were grown for five days. Images were captured using the Invitrogen EVOS M7000 Imaging System. Scale bar = 650 µm.

Access full white paper

Recent publications using HPLM

Cancer
Inmunology
Metabolism
Other research

Title	Cell type(s)	Research area(s)	Assay type
CRISPR screens in physiologic medium reveal conditionally essential genes in human cells	K562, MOLM-13, SUDHL4, and NOMO1	Cancer biology	CRISPR, RNA-seq, metabolite profiling, enzyme activity
De novo pyrimidine synthesis is a targetable vulnerability in IDH mutant glioma	BT054 oligodendroglioma cells	Cancer biology	transduction, drug screening, metabolite analysis, mass spec, isotope tracing, western blotting
Early reduction of glucose consumption is a biomarker of kinase inhibitor efficacy which can be reversed with GLUT1 overexpression in lung cancer cells	PC9, H1229, H3122	Cancer biology	growth kinetics, drug screening, xenograft, transduction
Activating mTOR mutations are detrimental in nutrient-poor conditions	primary mouse embryonic fibroblasts	Cancer biology	growth kinetics
Therapeutic efficacy of RAS inhibitor trametinib using a juvenile myelomonocytic leukemia patient-derived xenograft model	juvenile myelomonocytic leukemia (JMML) cells	Immuno-oncology	western blot

Title	Cell type(s)	Research area(s)	Assay type
Human Plasma-like Medium improves T lymphocyte activation	human and mouse T cells	Immunology	transcriptome analysis, NGS
MTHFD2 is a metabolic checkpoint controlling effector and regulatory T cell fate and function	primary human T cells	Immunology	T cell activation, metabolomics, mass spec, next-gen bisulfite sequencing, IHC, flow cytometry
Single-cell profiling of the antigen-specific response to BNT162b2 SARS-CoV-2 RNA vaccine	human PBMC	Immunology	LIBRA-seq, single-cell RNA-seq, flow cytometry
Plasmacytoid dendritic cell activation is dependent on coordinated expression of distinct amino acid transporters	plasmacytoid dendritic cells isolated from PBMCs	Immunology	ELISA, flow cytometry, isotope tracing, single-cell RNA-seq, CHIP-seq, IHC
mTOR regulation of metabolism limits LPS-induced monocyte inflammatory and procoagulant responses	human PBMC	Immunology	ELISA, flow cytometry, metabolite profiling, isotope labelling, RNA-seq, ChIP
Obesity and inflammation influence pharmacokinetic profiles of PEG-based nanoparticles	Sk-Hep1 human liver adenocarcinoma cells, Tp1 cells	Immunology, Obesity	fluorescence imaging, flow cytometry
Suppression of CEBPδ recovers exhaustion in anti-metastatic immune cells	mouse NK cells	Immunology	siRNA mediated knockdown, tumoricidal assay

Title	Cell type(s)	Research area(s)	Assay type
Physiologic medium rewires cellular metabolism and reveals uric acid as an endogenous inhibitor of UMP synthase	K562, KMS12BM, NOMO1, P12-Ichikawa, SEM, SUDHL4, 786-0, A549, MCF7, SW620, BJ, CLF-PED-015T, primary acute myeloid leukemia cells	Cancer metabolism	growth kinetics, metabolite quantification and analysis, cholesterol quantification, oxygen consumption
Inhibiting both proline biosynthesis and lipogenesis synergistically suppresses tumor growth	HeLa, MDA-MB-231, MCF-7, A549, HepG2, 8133, SKOV3, and mouse 4T1 cells	Cancer metabolism	electron production in metabolism, metabolite isotope tracing, RNA-seq
ZBTB1 regulates asparagine synthesis and leukemia cell response to L-asparaginase	CUTLL1, SUPT1, MOLM-13	Cancer metabolism	metabolite profiling, isotope tracing, ATAC-seq, mass spec
Lineage-specific silencing of PSAT1 induces serine auxotrophy and sensitivity to dietary serine starvation in luminal breast tumors	HCC1806, SUM149, BT549, HCC1937, HCC70, BT20, MCF7, MDA-MB-453, ZR75-1, EFM19, HCC1500, T47D	Cancer metabolism	metabolite analysis, cell proliferation, mass spec
Mitochondrial NADP+ is essential for proline biosynthesis during cell growth	HEK293E, HeLa, K562	Cancer metabolism	CRISPR, mitochondria isolation, spheroid growth, cell cycle, metabolic flux analysis, mass spec
Metabolic perturbations sensitize triple-negative breast cancers to apoptosis induced by BH3 mimetics	HCC1143, HCC1937, MDA-MB-231, MDA-MB-468	Cancer metabolism	metabolite analysis, BH3 profiling, qPCR
p53-mediated AKT and mTOR inhibition requires RFX7 and DDIT4 and depends on nutrient abundance	U2OS, HCT116	Cancer metabolism	transfection, ChIP, western blotting
Increased mitochondrial proline metabolism sustains proliferation and survival of colorectal cancer cells	RKO	Cancer metabolism	transfection, cell proliferation, apoptosis, mass spec
Human Plasma-Like Media fine tune mitochondrial function and alter drug sensitivity in cancer cell lines	SW620, MCF7, A375	Cancer metabolism	cell proliferation, respirometry
Human acute leukemia utilizes branched-chain amino acid catabolism to maintain stemness through regulating PRC2 function	human primary AML and ALL cells	Cancer metabolism	metabolome analysis, flow cytometry, transplantation, RNA microarray, ChIP-seq, cell cycle, isotope tracing
Methionine metabolism controls the B cell EBV epigenome and viral latency	P3HR-1 Burkitt’s lymphoma cell	Cancer metabolism	CRISPR, MeDIP, ChIP, RNA-seq, mouse xenograft, metabolite profiling, IHC
Elevated transferrin receptor impairs T cell metabolism and function in systemic lupus erythematosus	mouse naïve T cells, patient-derived T cells	Cancer metabolism	CRISPR, RNA-seq, mass spec, IHC
Formate overflow drives toxic folate trapping in MTHFD1 inhibited cancer cells	SW620 (CLL-227), HCT116 (CLL-247), MDA-MB-468 (HTB-132) and MDA-MB-231 (HTB-26) cells	Cancer metabolism	drug affinity responsive target stability (DARTS) assay, cellular thermal shift assay (CETSA), stable isotope tracing, mass spec
Canagliflozin impairs T cell effector function via metabolic suppression in autoimmunity	human PBMC-derived T cells	T cell metabolism	ELISA, flow cytometry, metabolic analysis by extracellular flux analyzer, western blot, stable isotope tracer analysis (SITA) by LC-MS, RNA-seq, proteomic analysis, mass spec
Iron is critical for mucosal-associated invariant T cell metabolism and effector functions	human PBMCs	T cell metabolism	transferrin uptake assay
Targeting fatty acid synthase in preclinical models of TNBC brain metastases synergizes with SN-38 and impairs invasion	MDA-MB-231	Cancer metabolism	drug screening, RNA expression
Hypoxanthine in the microenvironment can enable thiopurine resistance in acute lymphoblastic leukemia	NALM-6, REH, SEM, CEM, Jurkat	Cancer metabolism	drug assay (viability readouts using Cell-Titer Glo, Caspase 3/7 assay)
Conditional lethality profiling reveals anticancer mechanisms of action and drug-nutrient interactions	K562, SEM, NOMO1, P12-Ichikawa	Cancer metabolism	drug screening
Glucose limitation protects cancer cells from apoptosis induced by pyrimidine restriction and replication inhibition	Jurkat, Raji, A549, HCT116, MDA-MB-468, HEK293FT	Cancer metabolism	apoptosis
Coordination between the eIF2 kinase GCN2 and p53 signaling supports purine metabolism and the progression of prostate cancer	Human prostate cancer tumoroids (TM00298)	Cancer metabolism	cell cycle, metabolomics, ribosome biogenesis, CRISPR screening

Title	Cell type(s)	Research area(s)	Assay type
Nickels and tines: the myth of nickel allergy in intracranial stents	not applicable	Medicine/surgery	nickel allergy test
Generation of induced pluripotent stem cell-derived beta-cells in blood amino acids-like medium	human iPSCs	Stem cell differentiation	qPCR
The development of a smart magnetic resonance imaging and chemical exchange saturation transfer contrast agent for the imaging of sulfatase activity	NA (proprietary ligand linked to gadolinium)	Imaging enzyme activity	fast field cycling NMR relaxometry, z-spectrum
Quantification of cell death and proliferation of patient-derived ovarian cancer organoids through 3D imaging and image analysis	patient-derived ovarian cancer organoids	Drug screening	high content imaging and analysis
A new physiological medium uncovers biochemical and cellular alterations in Lesch-Nyhan disease fibroblasts	primary human skin fibroblasts from healthy individuals and those with Lesch-Nyhan disease	Disease biology	scratch assay, qPCR, mitochondrial respiration
Vitamin A resolves lineage plasticity to orchestrate stem cell lineage choices	mouse hair follicle stem cells	Stem cell differentiation	ATAC-seq, RNA-seq, CUT&RUN (CNR)
Human plasma-like medium (HPLM) induces Cryptococcus neoformans in vivo cell morphologies	Cryptococcus neoformans (C. neoformans)	Host-pathogen interaction	flow cytometry
CRISPR-based functional profiling of the Toxoplasma gondii genome during acute murine infection	toxoplasma gondii	Host-pathogen interaction	genome wide gene survey
Engineered myovascular tissues for studies of endothelial/satellite cell interactions	Primary human myogenic cells, primary Human endothelial progenitor cells	Tissue engineering	cell-cell interaction, cell differentiation

Order Gibco HPLM

Item	Cat. No
2-hydroxybutyric acid (available through Alfa Aesar)	Alfa Aesar A18636-03
Fetal bovine serum, dialyzed, US origin	26400044
6-Well Plate, TC Surface, Pack of 1<	140675
96-Well Plate, TC Surface, Pack of 1	167008
150 mm EasYDish, TC Surface, Pack of 10	150468
DPBS, no calcium, no magnesium	14190144
TrypLE Express Enzyme (1X), no phenol red	12604013
Trypsin-EDTA, 0.05%, phenol red	25300054
Trypan blue solution, 0.4%	15250061
Countess 3 Automated Cell Counter	AMQAX2000

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