Search Thermo Fisher Scientific
Search Thermo Fisher Scientific
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:
Order now View data Explore publications Watch webinar Request sample
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: “The composition of human plasma differs from conventional media, and we hypothesized that such differences could impact immune cell physiology. Here, we showed that relative to the medium typically used to culture lymphocytes (RPMI), a physiologic medium (human plasma-like medium; HPLM) induced markedly different transcriptional responses in human primary T cells and in addition, improved their activation upon antigen stimulation. We found that this medium-dependent effect on T cell activation is linked to Ca2+, which is six-fold higher in HPLM than in RPMI. Thus, a medium that more closely resembles human plasma has striking effects on T cell biology, further demonstrates that medium composition can profoundly affect experimental results, and broadly suggests that physiologic media may offer a valuable way to study cultured immune cells."
Reproduced with permission from: Leney-Greene MA, Boddapati AK, Su HC et al. (2020) Human plasma-like medium improves T lymphocyte activation. iScience 23:100759. doi: 10.1016/j.isci.2019.100759
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 |
“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
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:
Summary
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.
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: “The composition of human plasma differs from conventional media, and we hypothesized that such differences could impact immune cell physiology. Here, we showed that relative to the medium typically used to culture lymphocytes (RPMI), a physiologic medium (human plasma-like medium; HPLM) induced markedly different transcriptional responses in human primary T cells and in addition, improved their activation upon antigen stimulation. We found that this medium-dependent effect on T cell activation is linked to Ca2+, which is six-fold higher in HPLM than in RPMI. Thus, a medium that more closely resembles human plasma has striking effects on T cell biology, further demonstrates that medium composition can profoundly affect experimental results, and broadly suggests that physiologic media may offer a valuable way to study cultured immune cells."
Reproduced with permission from: Leney-Greene MA, Boddapati AK, Su HC et al. (2020) Human plasma-like medium improves T lymphocyte activation. iScience 23:100759. doi: 10.1016/j.isci.2019.100759
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 |
“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
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:
Summary
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.
Human Plasma-like Medium Improves T Lymphocyte Activation
Leney-Greene MA, Boddapati AK, Su HC, Cantor JR, Lenardo MJ. iScience. 2020;23(1):100759.
MTHFD2 is a Metabolic Checkpoint Controlling Effector and Regulatory T Cell Fate and Function
Sugiura A, Andrejeva G, Voss K, Heintzman DR, Beier KL, Wolf MM, Greenwood D, Ye X, Shahi SK, Freedman SN, Cameron AM, Foerch P, Bourne T, Xu X, Garcia-Canaveras JC, Mangalam AK, Rabinowitz JD, Rathmell JC. Immunity 2022 Jan 11;55(1):65-81.e9.
Plasmacytoid dendritic cell activation is dependent on coordinated expression of distinct amino acid transporters
Grzes KM, Sanin DE, Kabat AM, ..., Fabri M, Pearce EL, Pearce EJ. Immunity 2021 Nov 9;54(11):2514-2530.e7.
Physiologic Medium Rewires Cellular Metabolism and Reveals Uric Acid as an Endogenous Inhibitor of UMP Synthase
Cantor JR, Abu-Remaileh M, Kanarek N, Freinkman E, Gao X, Louissaint Jr A, Lewis CA, Sabatini DM. Cell 2017 Apr 6;169(2):258-272.
CRISPR screens in physiologic medium reveal conditionally essential genes in human cells
Rossiter NJ, Huggler KS, Adelmann CH, Keys HR, Soens RW, Sabatini DM, Cantor JR. Cell Metab 2021;S1550-4131(21)00061-9.
Lineage-Specific Silencing of PSAT1 Induces Serine Auxotrophy and Sensitivity to Dietary Serine Starvation in Luminal Breast Tumors
Choi BH, Conger KO, Selfors LM, Coloff JL. Cell Rep 2022 Jan 18;38(3):110278.
Inhibiting both proline biosynthesis and lipogenesis synergistically suppresses tumor growth
Liu M, Wang Y, Yang C, Ruan Y, Bai C, Chu Q, Cui Y, Chen C, Ying G, Li B. J Exp Med 2020 Mar 2;217(3):e20191226.
Systematic alteration of in vitro metabolic environments reveals empirical growth relationships in cancer cell phenotypes
Kochanowski K, Sander T, Link H, Chang J, Altschuler SJ, Wu LF. Cell Reports 2021 Jan 19;34,108647.
Mitochondrial NADP(+) is essential for proline biosynthesis during cell growth
Tran DH, Kesavan R, Rion H, Soflaee MH, Solmonson A, Bezwada D, Vu HS, Cai F, Phillips JA, DeBerardinis RJ, Hoxhaj G. Nat Metab 2021 Apr;3(4):571–585.
Human Plasma-like Medium Improves T Lymphocyte Activation
Leney-Greene MA, Boddapati AK, Su HC, Cantor JR, Lenardo MJ. iScience. 2020;23(1):100759.
MTHFD2 is a Metabolic Checkpoint Controlling Effector and Regulatory T Cell Fate and Function
Sugiura A, Andrejeva G, Voss K, Heintzman DR, Beier KL, Wolf MM, Greenwood D, Ye X, Shahi SK, Freedman SN, Cameron AM, Foerch P, Bourne T, Xu X, Garcia-Canaveras JC, Mangalam AK, Rabinowitz JD, Rathmell JC. Immunity 2022 Jan 11;55(1):65-81.e9.
Plasmacytoid dendritic cell activation is dependent on coordinated expression of distinct amino acid transporters
Grzes KM, Sanin DE, Kabat AM, ..., Fabri M, Pearce EL, Pearce EJ. Immunity 2021 Nov 9;54(11):2514-2530.e7.
Physiologic Medium Rewires Cellular Metabolism and Reveals Uric Acid as an Endogenous Inhibitor of UMP Synthase
Cantor JR, Abu-Remaileh M, Kanarek N, Freinkman E, Gao X, Louissaint Jr A, Lewis CA, Sabatini DM. Cell 2017 Apr 6;169(2):258-272.
CRISPR screens in physiologic medium reveal conditionally essential genes in human cells
Rossiter NJ, Huggler KS, Adelmann CH, Keys HR, Soens RW, Sabatini DM, Cantor JR. Cell Metab 2021;S1550-4131(21)00061-9.
Lineage-Specific Silencing of PSAT1 Induces Serine Auxotrophy and Sensitivity to Dietary Serine Starvation in Luminal Breast Tumors
Choi BH, Conger KO, Selfors LM, Coloff JL. Cell Rep 2022 Jan 18;38(3):110278.
Inhibiting both proline biosynthesis and lipogenesis synergistically suppresses tumor growth
Liu M, Wang Y, Yang C, Ruan Y, Bai C, Chu Q, Cui Y, Chen C, Ying G, Li B. J Exp Med 2020 Mar 2;217(3):e20191226.
Systematic alteration of in vitro metabolic environments reveals empirical growth relationships in cancer cell phenotypes
Kochanowski K, Sander T, Link H, Chang J, Altschuler SJ, Wu LF. Cell Reports 2021 Jan 19;34,108647.
Mitochondrial NADP(+) is essential for proline biosynthesis during cell growth
Tran DH, Kesavan R, Rion H, Soflaee MH, Solmonson A, Bezwada D, Vu HS, Cai F, Phillips JA, DeBerardinis RJ, Hoxhaj G. Nat Metab 2021 Apr;3(4):571–585.
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 |
Gibco cell culture products are manufactured in facilities compliant with current good manufacturing practices (GMP) and adhere to a robust quality management system, helping to ensure the consistency, reliability, and high quality you can rely on.
The 1 L Gibco bottle uses 38% less source material (32% less material for the 500 mL bottle) than comparable media bottles on the market today. This helps reduce the consumption of non-renewable resources, helping shrink your footprint.
Download the Green Fact Sheet
Learn more about sustainable solutions
Cell Culture & Transfection Learning Center
Access cell culture and transfection educational resources for better experiment planning and execution.
Gibco Cell Culture Basics
Learn the fundamentals of cell culture for achieving consistent results, including laboratory setup, safety, and aseptic techniques.
Media Formulation Tool
Find the right Gibco media formulation for DMEM, DMEM/F-12, MEM, and RPMI-1640 media.
Cell Culture & Transfection Support Center
Find technical support recommendations for your cell culture and transfection workflows, including tips for experimental setup and in-depth troubleshooting help.
Need technical support? Contact our expert team for technical and application support of Laboratory Products.
For Research Use Only. Not for use in diagnostic procedures.