It’s hard to believe barefacedtruth.com just celebrated its 10 year anniversary. Time does fly when you’re having fun! Your hosts are especially pleased that our readers have enjoyed our message and commitment to making high-science concepts of skincare understandable to people of all educational and cultural backgrounds. Questions from our readers clearly demonstrate that interest and have at times challenged us to delve deeply into the medical literature to answer complex “deep science” questions. We are impressed and proud of our readership.
We trust our mutual dialogue has furthered consumer understanding of an often difficult to understand product category. We hope our mission to “inform, educate and entertain” has been honored. Concerned and chagrined at the tendency of the skincare field to use “smoke and mirrors”, and all-to-often absurd pseudo-science claims to sell products to unsophisticated consumers, we pulled back the curtain more than once to expose outright (IOHO) subterfuge. We blew the whistle on several products about which we had serious ethical or scientific concerns. Some stalled, crashed and burned. Others howled “foul” and hired attorneys. We were undeterred and persevered in our mission, nonetheless. We’re glad we did.
It’s no secret your hosts, DrJohn and DrGeorge, have also been active this past decade in the formulating and marketing of what we consider “best-in-class” high-science products for skincare, hair regrowth, and women’s intimate health. Our company has grown exponentially and now has distribution in many foreign countries, with more planned in the near future. Our products are used in thousands of medical and esthetic practices every day, often as pro-healing and anti-inflammatory adjuvants to a variety of non or minimally invasive medical procedures. More products are in the pipeline.
Below is posted a document recently prepared for our sales team that answers questions that have been asked by physicians and estheticians considering adopting Cellese products into their practices. We think our readership will find it interesting, especially those who date back to our earlier years when we were busy explaining the field of human stem cells, conditioned media, and secretome composition and how they apply and impact skin health, procedure recovery, and anti-aging.
BONE MARROW MESENCHYMAL STEM CELLS: THE BODY’S EMISSARY CELL FOR TISSUE REPAIR & REGENERATION
At its founding, Cellese physician/scientists committed to creating best-in-class high-science antiaging skincare products. Because skin aging results from cumulated injuries and related inflammation, our flagship products – AnteAGE and AnteAGE MD Serum & Accelerator – were formulated using “biomimicry” to emulate how the human body naturally heals and repairs injuries in skin (and other tissues). Our decision to focus on exploiting the unique properties of bone marrow mesenchymal stem cells in our formulation was well considered and based on abundant published scientific evidence.
Bone marrow mesenchymal stem cells (“BMMSCs”) are unique among the several populations of stem cells within the human body. BMMSCs are migratory and circulate within the blood to reach all tissues. When they encounter injury, they are tasked with orchestrating the symphony of healing cascades that repairs damaged tissue. Paramount among these are modulation and amelioration of inflammation, which is a major driver of skin aging.
Of note, comparison of the anti-inflammatory potency of BMMSCs compared to adipose-derived mesenchymal stem cells (“ASCs”) was clearly demonstrated in a rodent model of endotoxemia:
“Bone marrow-derived mesenchymal stromal cells (BMMSCs) are a cell population of intense exploration for therapeutic use in inflammatory diseases. Secreted factors released by BMSCs are responsible for the resolution of inflammation in several pre-clinical models. [….] Proteomic profiling of conditioned media revealed that BMMSCs express significantly higher levels of sVEGFR1 and sTNFR1, two soluble cytokine receptors with known therapeutic activity in sepsis. In a prophylactic study of endotoxin-induced inflammation in mice, we observed that BMMSCs secreted factors provided a greater survival benefit and tissue protection of endotoxemic mice compared to ASCs.”
BMMSCs influence local physiologic processes through secretion of cell-to-cell messenger molecules, collectively referred to as their secretome. Abundant evidence supports that stem cell therapies based on use of the secretome (conditioned media, exosomes, etc.) emulates, and is in important ways superior to therapies using stem cells per se. The Cellese products containing conditioned media derived from laboratory culture of BMMSCs exploit these benefits.
Cellese scientists are currently developing techniques and methods to expand BMMSC secretome delivery to include exosomes, tiny vesicles released from cell membranes that contain high concentrations of DNA, mRNA, miRNA, proteins (including cytokines and growth factors), lipids and other bioactive molecules. Published evidence strongly suggests delivery of BMMSC-derived exosomes to iatrogenically or otherwise injured skin will produce substantial pro-healing and anti-inflammatory clinical benefits. (Our scientific inquiry into exosomes will also examine benefits offered and unique to other types of stem cells. Proven advantages, if any, will drive decisions as to preferred stem cell population to exploit for our exosome projects.)
A 2017 article in the International Journal of Molecular Science summarized that exosomes derived from bone marrow mesenchymal stem cells have proven of benefit in at least one clinically relevant parameter in the following diseases and conditions:
- Reduced infarct size in myocardial ischemia / reperfusion injury
- Renoprotection in acute kidney disease
- Reduced pulmonary inflammation in acute lung injury
- Attenuation of injury in acute pancreatitis
- Improved hepatic regeneration in liver injury
- Suppression of tumor progression and angiogenesis
- Promoting breast cancer cell dormancy in metastatic niche
- Rescue of bone marrow function in lupus
- Prevention of abnormal neurogenesis and memory deficit in epilepsy
- Increased tumor death in glioblastoma
- Inhibited Kaposi’s sarcoma growth
This contrasts with more limited improvements seen with umbilical cord perivascular, adipose-derived, Wharton’s Jelly-derived, and periodontal ligament-derived stem cells. Improvements with exosomes from these other stem cell population number 4, 3, 1, and 1 respectively.
References:
Andrzejewska A, Lukomska B, Janowski M. Concise Review: Mesenchymal Stem Cells: From Roots to Boost. Stem Cells. 2019 Jul;37(7):855-864. doi: 10.1002/stem.3016. Epub 2019 Apr 30. PMID: 30977255; PMCID: PMC6658105.
Fu X, Liu G, Halim A, Ju Y, Luo Q, Song AG. Mesenchymal Stem Cell Migration and Tissue Repair. Cells. 2019 Jul 28;8(8):784. doi: 10.3390/cells8080784. PMID: 31357692; PMCID: PMC6721499.
Harrell CR, Fellabaum C, Jovicic N, Djonov V, Arsenijevic N, Volarevic V. Molecular Mechanisms Responsible for Therapeutic Potential of Mesenchymal Stem Cell-Derived Secretome. Cells. 2019 May 16;8(5):467. doi: 10.3390/cells8050467. PMID: 31100966; PMCID: PMC6562906.
Elman JS, Li M, Wang F, Gimble JM, Parekkadan B. A comparison of adipose and bone marrow-derived mesenchymal stromal cell secreted factors in the treatment of systemic inflammation. J Inflamm (Lond). 2014 Jan 7;11(1):1. doi: 10.1186/1476-9255-11-1. PMID: 24397734; PMCID: PMC3895743
Vizoso FJ, Eiro N, Cid S, Schneider J, Perez-Fernandez R. Mesenchymal Stem Cell Secretome: Toward Cell-Free Therapeutic Strategies in Regenerative Medicine. Int J Mol Sci. 2017 Aug 25;18(9):1852. doi: 10.3390/ijms18091852. PMID: 28841158; PMCID: PMC5618501.
Gwam C, Mohammed N, Ma X. Stem cell secretome, regeneration, and clinical translation: a narrative review. Ann Transl Med. 2021 Jan;9(1):70. doi: 10.21037/atm-20-5030. PMID: 33553363; PMCID: PMC7859812.
Vizoso FJ, Eiro N, Cid S, Schneider J, Perez-Fernandez R. Mesenchymal Stem Cell Secretome: Toward Cell-Free Therapeutic Strategies in Regenerative Medicine. Int J Mol Sci. 2017 Aug 25;18(9):1852. doi: 10.3390/ijms18091852. PMID: 28841158; PMCID: PMC5618501.
ADDRESSING RECENT QUESTIONS POSED CONCERNING CONDITIONED MEDIA (“CM”)
- Regarding a CM secretome, does the source of stem cells matter, or only the number of cells in culture and laboratory growth conditions?
The stem cell niche from which each type of stem cells is derived reflects their inherent unique genetic destiny, i.e., their tissue of origin. It therefore follows that the secretomes they produce in culture differ for the same reason (genetic differences), although there may be substantial commonality of bioactive molecules. This has been studied and validated using mass spectrometry and bioinformatics. Each mesenchymal stem cell niche produces a secretome with its own distinct characteristics, although they may share specific properties e.g., cell migration and regulation of programmed cell death.
Production of CM involves harvesting post-culture nutrient broth and replenishing it with fresh broth during the process of multiple “passages”. A passage occurs each time a culture is subcultured, i.e., harvested and reseeded into multiple “daughter” cell culture flasks. Stem cell culture passages are required whenever stem cells (which are by nature adherent and grow in single layers) proliferate to become 60% to 80% confluent. Cell confluence is a major inhibitory stimulus to further cell proliferation. Arrested growth dramatically reduces or terminates continued secretome production. Implicitly, one can assume more cells yield more secretome content and that some cells may be more proliferative producers than others. Larger surface area flasks provide more “room to grow” prior to confluence induced proliferation arrest. Laboratory conditions can be modified in a variety of ways to influence secretome composition.
The quote below is taken from a 2018 review article in Frontiers in Immunology (citation below).
“Although MSCs have an innate potential to induce and/or contribute to regeneration, this potential is now known to be greatly influenced by diverse extrinsic factors such as the tissue source of the MSCs, the health status and age of the MSCs donor, the batch/lot of serum used for the in vitro culture of the MSCs, passage number, oxygen concentration, and the presence/absence of pro-inflammatory bioactive molecules within the culture milieu. Thus, in vitro preconditioning of MSCs with a variety of different factors has been explored to enhance the therapeutic capacity/potential of MSCs, which included: 3D culture, pharmacological compounds, inflammatory cytokines and hypoxia.”
Colter DC, Class R, DiGirolamo CM, Prockop DJ. Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow. Proc Natl Acad Sci U S A. 2000 Mar 28;97(7):3213-8. doi: 10.1073/pnas.97.7.3213. PMID: 10725391; PMCID: PMC16218.
Ferreira JR, Teixeira GQ, Santos SG, Barbosa MA, Almeida-Porada G, Gonçalves RM. Mesenchymal Stromal Cell Secretome: Influencing Therapeutic Potential by Cellular Pre-conditioning. Front Immunol. 2018 Dec 4;9:2837. doi: 10.3389/fimmu.2018.02837. PMID: 30564236; PMCID: PMC6288292.
- Do stem cells from bone marrow and fat excrete the same secretome? In the same amount?
Although both are categorized as mesenchymal stem cells because of their origin within the mesenchyme, the embryologically primitive ground substance matrix of the mesoderm, bone marrow and fat stem cells have different genetic “blueprints” that control their characteristics and function. It is therefore logical that their secretomes have both similarities and differences.
A 2015 research article in Stem Cell Research and Therapy disclosed that:
“Primary ASCs and BMSCs have highly similar small RNA expression profiles dominated by miRNAs and snoRNAs (together 64-71 %), of which 150-200 miRNAs are present at physiological levels [….] While ASC and BMSC exosomes are similar in RNA class distribution and composition, we observed striking differences in the sorting of evolutionary conserved tRNA species that seems associated with the differentiation status of MSCs [….]”
Baglio SR, Rooijers K, Koppers-Lalic D, Verweij FJ, Pérez Lanzón M, Zini N, Naaijkens B, Perut F, Niessen HW, Baldini N, Pegtel DM. Human bone marrow- and adipose-mesenchymal stem cells secrete exosomes enriched in distinctive miRNA and tRNA species. Stem Cell Res Ther. 2015 Jul 1;6(1):127. doi: 10.1186/s13287-015-0116-z. PMID: 26129847; PMCID: PMC4529699.
A 2019 review article in Cytokine and Growth Factor Reviews observed:
“MSCs harvested from numerous anatomical locations including the bone marrow, adipose tissue, Wharton’s jelly of the umbilical cord etc., display similar immunophenotypic profiles. However, there is a large body of evidence showing that MSCs secrete a variety of biologically active molecules such as growth factors, chemokines, and cytokines. Despite the similarity in their immunophenotype, the secretome of MSCs appears to vary significantly, depending on the age of the host and niches where the cells reside. Thus, by implication, proteomics-based profiling suggests that the therapeutic potential of the different MSC populations must also be different.”
L PK, Kandoi S, Misra R, S V, K R, Verma RS. The mesenchymal stem cell secretome: A new paradigm towards cell-free therapeutic mode in regenerative medicine. Cytokine Growth Factor Rev. 2019 Apr;46:1-9. doi: 10.1016/j.cytogfr.2019.04.002. Epub 2019 Apr 2. PMID: 30954374.
- Do all stem cells proliferate at the same rate in culture?
A large variety of factors influence the rate of proliferation of stem cells in culture so the short answer is “no”, the rate of proliferation can vary significantly. (There are also positive and negative influences in stem cell proliferation in vivo.) In addition to donor age and gender, a variety of factors during the culture process have significant impact on cell proliferation.
According to manufacturers of cell culture devices, factors that affect cell proliferation are:
- Temperature
- Osmotic pressure
- Oxygen, carbon dioxide, and pH values
- Non-toxic and sterile environments
- Radiation and ultrasound influences, including visible light and UV light
- Cell seeding density
- Container rotation speed (if in a rotating culture container)
Fossett E, Khan WS, Longo UG, Smitham PJ. Effect of age and gender on cell proliferation and cell surface characterization of synovial fat pad derived mesenchymal stem cells. J Orthop Res. 2012 Jul;30(7):1013-8. doi: 10.1002/jor.22057. Epub 2012 Jan 6. PMID: 22228598.
https://www.zhichushakers.com/7-factors-that-affect-cell-growth.html
- How is BMMSC condition media different from that of other cultured stem cells?
What is in it?
Recognized characteristics of BMMSCs in abundant published peer-reviewed studies, and important in the selection of its CM for use in Cellese products, are its immunomodulatory, anti-inflammatory, and pro-healing properties – the reasons BMMSCs were selected to source our CM ingredients.
These properties in our products were anecdotally confirmed by early purchasers of AnteAGE and AnteAGE MD Serum (during 2012-2014), who described more rapid than usual healing of abrasions, thermal injury, and sun burns. Several also reported they did not develop the PIH they had seen with healing of previous injuries. In 2014, the president of an American laser distributor performed a split-face study on himself comparing AnteAGE Serum in aftercare following fractional CO2 laser resurfacing. The study side was fully recovered in less than four days, whereas the untreated side took over six days. He reported less edema, erythema, and pain on the study side. His company recommends use of AnteAGE products as part of its post-procedure protocol.
Globally, several thousand medical and esthetic practices use Cellese products routinely as adjuvants in a variety of non and minimally invasive esthetic skin procedures. Many also report patients experience significant clinical benefit when certain products are used “off label” for intradermal injection. After a decade of producing and distributing products containing BMMSC CM, user and practitioner examples of pro-healing anti-inflammatory benefit using Cellese products are too numerous to count.
In the world’s literature, the number of studies exploring the clinical benefits of BMMSCs and derived CM outnumber studies of mesenchymal stem cells from other niches severalfold. One example is the use of BMMSC CM as a treatment for allergic rhinitis.
From a 2018 article about this indication published in the journal, Medical Hypothesis:
“Bone marrow mesenchymal stem cells (BMSCs) are a population of adult stem cells with multipotential differentiation capability, low immunogenicity, and immunoregulatory effects. The unique immunoregulatory properties of BMSCs make them hold great promise in the treatment of chronic inflammation and immune disorders through a paracrine mechanism of anti-inflammatory and anti-allergic effects.”
Zou W, Liu G, Zhang J. Secretome from bone marrow mesenchymal stem cells: A promising, cell-free therapy for allergic rhinitis. Med Hypotheses. 2018 Dec;121:124-126. doi: 10.1016/j.mehy.2018.09.016. Epub 2018 Sep 10. PMID: 30396464.
BMMSCs and derived CM are also being studied for possible clinical benefit in many other indications. A 2020 article in the International Journal of Molecular Science summarized the number of trials underway at that time.
The major endocrine factors secreted by bone marrow mesenchymal stem cells are tabulated below:
Chu DT, Phuong TNT, Tien NLB, Tran DK, Thanh VV, Quang TL, Truong DT, Pham VH, Ngoc VTN, Chu-Dinh T, Kushekhar K. An Update on the Progress of Isolation, Culture, Storage, and Clinical Application of Human Bone Marrow Mesenchymal Stem/Stromal Cells. Int J Mol Sci. 2020 Jan 21;21(3):708. doi: 10.3390/ijms21030708. PMID: 31973182; PMCID: PMC7037097.
BMMSCs exist in relatively small numbers within the bone marrow and obtaining them requires invasive procedures, although small quantities of marrow are sufficient to enable in vitro culture. This procedure can be performed easily on alert awake donors using local anesthetic. Large donations, such as are required for bone marrow transplantation, is more involved with much more marrow volume extracted and requires general anesthesia.
Extensive research over the past decade has focused on mesenchymal stem cells from other tissue niches (adipose, dental pulp, umbilical cord perivascular, placenta, Whartan’s Jelly, etc.) Similarities and differences in the properties of the CM derived from each type of stem cell have been explored. Many of the same bioactive molecules are secreted by stem cells from differing niches. There are important differences, however. It is the “balance” between bioactive molecules, especially the pro and anti-inflammatory balance, that was a major consideration when Cellese scientists selected BMMSCs as their niche of choice. A decade of experience has proven that to have been a wise and appropriate choice for anti-aging skincare products, deliberately formulated to emphasize the anti-inflammatory potency of BMMSC CM.
First User Experience: On July 1, 2011, a principal of Cellese, who for many years used daily oral doxycycline and topical metrogel to control erythematotelangiectatic rosacea, discontinued his pharmaceuticals and never again restarted them. He was the first person to trial the new AnteAGE Serum formula. More than a decade later using only Cellese products, he has not had another flareup – despite complete disregard for the several common triggers known to cause flareups.
Extracted comments from published literature that compares stem cell niche properties.
“Extracellular vesicles (EVs) derived from mesenchymal stem cells isolated from both bone marrow (BMSCs) and adipose tissue (ADSCs) show potential therapeutic effects[….] but exert different biological effect. [….] both ADSC and BMSC-EVs demonstrated beneficial effect on cells involved in skin wound healing such as fibroblasts, keratinocytes and endothelial cells, but through different cellular processes. Consistent with the bioinformatic analyses, BMSC-EVs were shown to mainly promote proliferation, whereas ADSC-EVs demonstrated a major effect on angiogenesis.”
Pomatto M, Gai C, Negro F, Cedrino M, Grange C, Ceccotti E, Togliatto G, Collino F, Tapparo M, Figliolini F, Lopatina T, Brizzi MF, Camussi G. Differential Therapeutic Effect of Extracellular Vesicles Derived by Bone Marrow and Adipose Mesenchymal Stem Cells on Wound Healing of Diabetic Ulcers and Correlation to Their Cargoes. Int J Mol Sci. 2021 Apr 8;22(8):3851. doi: 10.3390/ijms22083851. PMID: 33917759; PMCID: PMC8068154.
“Studies published in recent years have shown that the secretome of hMSCs isolated from different tissue sources may present significant variation. The present work performed a comparative proteomic-based analysis through mass spectrometry on the secretome of hMSCs derived from bone marrow (BMSCs), adipose tissue (ASCs), and human umbilical cord perivascular cells (HUCPVCs). The results revealed that BMSCs, ASCs, and HUCPVCs differed in their secretion of neurotrophic, neurogenic, axon guidance, axon growth, and neurodifferentiative proteins, as well as proteins with neuroprotective actions against oxidative stress, apoptosis, and excitotoxicity, which have been shown to be involved in several CNS disorder/injury processes. Although important changes were observed within the secretome of the cell populations that were analyzed, all cell populations shared the capability of secreting important neuroregulatory molecules.”
Pires AO, Mendes-Pinheiro B, Teixeira FG, Anjo SI, Ribeiro-Samy S, Gomes ED, Serra SC, Silva NA, Manadas B, Sousa N, Salgado AJ. Unveiling the Differences of Secretome of Human Bone Marrow Mesenchymal Stem Cells, Adipose Tissue-Derived Stem Cells, and Human Umbilical Cord Perivascular Cells: A Proteomic Analysis. Stem Cells Dev. 2016 Jul 15;25(14):1073-83. doi: 10.1089/scd.2016.0048. Epub 2016 Jun 27. PMID: 27226274.
“Mesenchymal stem cells (MSCs) have the potential to be a viable therapy against various diseases due to their paracrine effects, such as secretion of immunomodulatory, trophic and protective factors. These cells are known to be distributed within various organs and tissues. Although they possess the same characteristics, MSCs from different sources are believed to have different secretion potentials and patterns, which may influence their therapeutic effects in disease environments. We characterized the protein secretome of adipose (AD), bone marrow (BM), placenta (PL), and Wharton’s jelly (WJ)-derived human MSCs by using conditioned media and analyzing the secretome by mass spectrometry and follow-up bioinformatics. Each MSC secretome profile had distinct characteristics depending on the source. However, the functional analyses of the secretome from different sources showed that they share similar characteristics, such as cell migration and negative regulation of programmed cell death, even though differences in the composition of the secretome exist.”
Brown C, McKee C, Bakshi S, Walker K, Hakman E, Halassy S, Svinarich D, Dodds R, Govind CK, Chaudhry GR. Mesenchymal stem cells: Cell therapy and regeneration potential. J Tissue Eng Regen Med. 2019 Sep;13(9):1738-1755. doi: 10.1002/term.2914. Epub 2019 Jul 25. PMID: 31216380.
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