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Gibco Human Plasma-like Medium (HPLM) is formulated to resemble the natural cellular environment found in the body, mimicking the metabolic profile of human plasma.
Widely used, synthetic cell culture media, including MEM, DMEM, RPMI 1640, and DMEM/F-12 contain glucose, amino acids, vitamins, and salts at concentrations that, in large part, do not reflect those found in human plasma. These media also lack additional plasma components needed to mimic the metabolic profile of human plasma. When studying cancer and other diseases, results with more physiological relevance will enable researchers to help improve their understanding of human function and illness.
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:
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 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.
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
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.
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).
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).
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.
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 |
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:
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
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.
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 |
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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