Pure Exosome Therapy

A 99.9% Pure Exosome Intravenous Therapy Processed from Cultured Supernatant of Deciduous Dental Pulp Stem Cells

What are Exosomes?

Exosomes refer to tiny vesicles, ranging from 50 to 150 nanometers in size, enclosed by a lipid membrane found within the body.

Exosomes contain microRNAs, messenger RNAs, DNA, proteins, and other molecules. Among these, microRNAs and messenger RNAs have been recently discovered to play a crucial role in cellular communication, enabling cells to exchange information with each other.

Utilizing the properties of exosomes like these has the potential to be effective in various fields, including aging care and beyond.

The potential of exosomes is significant, as they impact the human body’s 37 trillion cells in various ways, playing a crucial role in cellular changes and the daily maintenance of life.

Currently, we are promoting exosome therapy derived from deciduous dental pulp stem cells and data analysis at affiliated medical facilities.

What is Pure Exo?

Pure Exo is a product made by purifying exosomes extracted from deciduous dental pulp stem cells with 99.9% purity. When using cultured supernatant, a substantial amount of it had to be administered to introduce exosomes into the body. However, with Pure Exo, it is possible to administer approximately 10 times the amount of exosomes found in cultured supernatant with only about 1/10 of the volume.

Cultured supernatant contains various growth factors, cytokines, and culture media in addition to exosomes. Since Pure Exo isolates only exosomes, it can deliver a large quantity of exosomes in a single administration without causing a cytokine storm.

SGF5ml contains 10 billion exosomes, but Pure Exo can provide approximately 100-200 billion exosomes in a single administration.

With SGF, it is possible to administer 5-20 cc (10-500 billion exosomes) at once, but for more than 100 billion exosomes, Pure Exo is required.

Recommended for individuals like this.

Expected Benefits

Treatment・Achievements Using Exosomes at R&L Clinic

・Prevention・treatment of ischemic diseases such as arteriosclerosis
・Improvement of vascular endothelial dysfunction
・Prevention and improvement of chronic internal medicine diseases such as diabetes
・Impact on telomere extension
【Treatment Method】Intravenous injection ・ intravenous drip

・Erectile Dysfunction (ED)
・Androgenetic Alopecia (AGA)
・Relief of pain in conditions like deformed elbow and knee joints
・Male infertility treatment
・Skin texture improvement
【Treatment Method】Local injection

・Improvement and prevention of dementia
・Improvement of side effects of stroke (such as hemiparesis)
・Alzheimer’s type dementia
・Mental disorders such as bipolar disorder
【Treatment Method】Nasal administration

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・Improvement in Vision

Improvement in Vision

PURE EXO 100（100 Billion Exosomes）	165,000 JPY
PURE EXO 1000（1 Trillion Exosomes）	1,100,000 JPY

※All prices include tax. Our clinic operates on a fee-for-service basis (not covered by public health insurance).

Exosome Structure

Exosomes are produced when membrane-bound vesicles, created by the inward budding of endosomes (a cellular mechanism for internalizing substances from the extracellular space), are released into the extracellular environment. Exosomes contain materials from the originating cell in their interior, and their surface is adorned with membrane proteins such as tetraspanins (CD9, CD63, CD81), integrins, and major histocompatibility complex (MHC) molecules. Furthermore, they contain numerous proteins internally related to multivesicular body formation (Tsg101, Alix) and heat shock proteins (HSP).

Exosomes serve as messengers for intercellular communication between cells

Exosomes are released from cells and circulate throughout the body via the bloodstream. During this process, the microRNAs and messenger RNAs contained within exosomes communicate with cells belonging to various organs in the body, influencing various actions. For example, they may signal, ‘Promote metabolism’ or ‘Create blood vessels.’

Such communication occurs not only between different types of cells but also between various organs. In these interactions, the presence of exosomes as messengers is crucial.

Traditionally, it was believed that all such communication occurred exclusively through the brain. However, research on exosomes has revealed that this is not the sole mechanism. For instance, when we overeat, our bodies naturally increase metabolism and energy consumption. The reason we can maintain good health in our daily lives is that our organs collaborate and communicate with each other, and exosomes play a vital role in these processes.

The intricate communication between cells and organs, facilitated by exosomes and other factors, ensures that each organ functions properly, contributing to our overall health.

Differences Between Stem Cell Culture Supernatant and Stem Cell Exosomes

“Stem cell culture supernatant and exosomes are not the same.”

For stem cell culture supernatant, it is crucial to confirm the manufacturing process and quality control. Quality should be assessed not merely based on the total number of particles but by verifying the purity and markers (CD9, CD63) of stem cell exosomes. Other particles aside from stem cell exosomes may potentially be harmful to the body.

Exosomes secreted by mesenchymal stem cells (MSCs) have been extensively researched by numerous scientists for their potential medical applications, and their therapeutic effects have been reported. Research papers have documented that exosomes derived from mesenchymal stem cells exhibit effects equivalent to or even greater than those of the stem cells themselves.

Expert Opin Biol Ther. 2016;16(4):489-506. doi: 10.1517/14712598.2016.1131976. Epub 2016 Jan 28.
Exosomes for repair, regeneration and rejuvenation Joydeep Basu, John W Ludlow

Promotion of Adipocyte Vascularization

In a study using mice, the comparison of the engraftment of adipose stem cell exosomes, adipose stem cells (processed), and adipose cells (tissues) in combination with physiological saline was conducted to investigate vascular neogenesis and engraftment levels. The results showed a significant difference between the group treated with adipose stem cell exosomes and adipose stem cells (processed) compared to the group with physiological saline alone. Furthermore, when comparing the group treated with exosomes alone to the group using adipose cells, the exosomes exhibited an equivalent level of engraftment as when using adipose cells. This suggests that exosomes have effects comparable to stem cell transplantation.

Repair, Regeneration, and Anti-Aging Effects of Mesenchymal Stem Cell Exosomes

Various studies have been conducted on the effects of mesenchymal stem cell exosomes in repairing, regenerating, and providing anti-aging care for cells and tissues, as evidenced by the following research papers. These studies have yielded numerous findings regarding the effectiveness of mesenchymal stem cell exosomes.

Expert Opin Biol Ther. 2016;16(4):489-506. doi: 10.1517/14712598.2016.1131976. Epub 2016 Jan 28.
Exosomes for repair, regeneration and rejuvenation　Joydeep Basu, John W Ludlow

3. Maguire G. Stem cell therapy without the cells. Commun Integr Biol. 2013;6:e26631. DOI:10.4161/cib.26631.

5. Caplan AI, Correa D. The MSC: an injury drugstore. Cell Stem Cell. 2011;9:11-15.

13. Valadi H, Ekstrom K, Bossios A, et al. Exosome mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nature Cell Biol. 2007;9:654-659. Demonstration of exosome-mediated communication between cells during development

14. Ratajczak J, Miekus K, Kucia M, et al. Embryonic stem cell derived micro-vesicles reprogram hematopoietic pro-genitors: evidence for horizontal transfer of mRNA and protein delivery. Leukemia. 2006;20:847-856.

19. Yanez-Mo M, Siljander PR, Andreu Z, et al. Biological proper-ties of extracellular vesicles and their physiological functions. J Extracell Vesicles. 2015;4. DOI:10.3402/jev.v4.27066.

32. Deregibus MC, Cantaluppi V, Calogero R, et al. Endothelial progenitor cell derived microvesicles activate an angiogenic program in endothelial cells by a horizon-tal transfer of mRNA. Blood. 2007;110:2440-2448.

40. Teixeira FG, Carvalho MM, Sousa N, et al. Mesenchymal stem cells secretome: a new paradigm for central ner-vous system regeneration. Cell Mol Life Sci. 2013;70:3871-3882.

41. Vishnubhatla I, Corteling R, Stevanato L, et al. The devel-opment of stem cell-derived exosomes as a cell-free regenerative medicine. J Circ Biomark. 2014;3:2. DOI:10.5772/58597.

43. Liang X, Ding Y, Zhang Y, et al. Paracrine mechanisms of mesenchymal stem cell-based therapy: current status and perspectives. Cell Transplant. 2013;9:1045-1059.

44. van Koppen A, Joles JA, van Balkom BW, et al. Human embryonic mesenchymal stem cell-derived conditioned medium rescues renal function in rats with established chronic kidney disease. PLoS ONE.

（？）Ionescu L, Byrne RN, van Haaften T, et al. Stem cell conditioned medium improves acute lung injury in mice: in vivo evidence of stem cell paracrine action. Am J Physiol Lung Cell Mol Physiol. 2012;303:L967–97.

45. Ratajczak MZ, Kucia M, Jadczyk T, et al. Pivotal role of paracrine effects in stem cell therapies in regenerative medicine: can we translate stem cell-secreted paracrine factors and microvesicles into better therapeutic strate-gies. Leukemia. 2012;26:1166-1173.

50. Osugi M, Katagiri W, Yoshimi R, et al. Conditioned media from mesenchymal stem cells enhanced bone regenera-tion in rat calvarial bone defects. Tissue Eng Part A. 2012;18:1479-1489.

51. Maumus M, Jorgensen C, Noel D. Mesenchymal stem cells in regenerative medicine applied to rheumatic dis-eases: role of secretome and exosomes. Biochimie. 2013;95:2229-2234.

52. Toh WS, Foldager CB, Pei M, et al. Advances in mesench-ymal stem cell-based strategies for cartilage repair and regeneration. Stem Cell Reviews and Reports. 2014;10:686-696.

53. Akyurekli C, Le Y, Richardson RB, et al. A systematic review of preclinical studies on the therapeutic potential of mesenchymal stromal cell-derived microvesicles. Stem Cell Rev. 2015;11:150-160. Review of key preclinical studies associated with mesenchymal stem cell (MSC)-sourced exosomes

55. Gatti S, Bruno S, Deregibus MC, et al. Microvesicles derived from human adult mesenchymal stem cells pro-tect against ischaemia reperfusion induced acute and chronic kidney injury. Nephrol Dial Transplant. 2011;26:1474-1483.

56. Wang Y, Zhang L, Li Y, et al. Exosomes/microvesicles from induced pluripotent stem cells deliver cardioprotec-tive miRNAs and prevent cardiomyocyte apoptosis in the ischemic myocardium. Int J Cardiol. 2015;192:61-69.

57. Kilpinen L, Impola U, Sankkila L, et al. Extracellular membrane vesicles from umbilical cord blood derived MSC protect against ischemic acute kidney injury, a feature that is lost after inflammatory conditioning. J Extracell Vesicles. 2013;10:3402.

60. Lamichhane TN, Sokic S, Schardt JS, et al. Emerging roles for extracellular vesicles in tissue engineering and regen-erative medicine. Tissue Eng Part B Rev. 2015;21:45-54. DOI:10.1089/ten.teb.2014.0300.

61. Sahoo S, Klychko E, Thorne T, et al. Exosomes from human CD34+ stem cells mediate their proangiogenic paracrine activity. Circ Res. 2011;109:724-728.

62. Lee C, Mitsialis SA, Aslam M, et al. Exosomes mediate the cytoprotective action of mesenchymal stromal cells on hypoxia induced pulmonary hypertension. Circulation. 2012;126:2601-2611.

63. Zhang J, Guan J, Niu X, et al. Exosomes released from human induced pluripotent stem cells derived MSCs facilitate cutaneous wound healing by promoting col-lagen synthesis and angiogenesis. J Trans Med. 2015;13:49. DOI:10.1186/s12967-015-0417-0.

64. Ibrahim AG, Cheng K, Marban E. Exosomes as critical agents of cardiac regeneration triggered by cell therapy. Stem Cell Reports. 2014;2:606-619.

71. Lee HK, Finniss S, Cazacu S, et al. Mesenchymal stem cells deliver exogenous miRNAs to neural cells and induce their differentiation and glutamate transporter expres-sion. Stem Cells Dev. 2014;23:2851-2861.

73. Zhuang X, Xiang X, Grizzle W, et al. Treatment of brain inflammatory diseases by delivering exosome-encapsulated anti-inflammatory drugs from the nasal region to the brain. Mol Ther. 2011;19:1769-1779. DOI:10.1038/mt.2011.164.

74. Xin H, Li Y, Liu Z, et al. Systematic administration of exo-somes released from mesenchymal stromal cells promote functional recovery and neurovascular plasticity after stroke in rats. J Cereb Blood Flow Metab. 2013;33:1711-1715.

75. Zhang Y, Chopp M, Meng Y, et al. Effect of exosomes derived from multipluripotent mesenchymal stromal cells on functional recovery and neurovascular plasticity in rats after traumatic brain injury. J Neurosurg. 2015;122:856-867.

76. Doeppner TR, Herz J, Gorgens A, et al. Extracellular vesi-cles improve post-stroke neuroregeneration and prevent postischemic immunosuppression. Stem Cells Transl Med. 2015;4:1131-1143.

77. Xin H, Li Y, Liu Z, et al. MiR-133b promotes neural plas-ticity and functional recovery after treatment of stroke with multipotent mesenchymal stromal cells in rats via transfer of exosome-enriched extracellular particles. Stem Cells. 2013;31:2737-2746.

78. Bruce A, Ilagan R, Guthrie K, et al. Selected renal cells modulate disease progression in rodent models of chronic kidney disease via NFκB and TGFβ1 pathways. Regen Med. 2015;10:815-839.

79. Fleury A, Martinez MC, Le Lay S. Extracellular vesicles as therapeutic tools in cardiovascular diseases. Front Immunol. 2014;5:370. DOI:10.3389/fimmu.2014.00370.

82. Vlassov AV, Magdaleno S, Setterquist R, et al. Exosomes: current knowledge of their composition, biological func-tions, and diagnostic and therapeutic potentials. Biochimica et Biophysica Acta. 2012;1820:940-948.

83. Kordelas L, Rebmann V, Ludwig A-K, et al. 2013. MSC-derived exosomes: a novel tool to treat therapy-refractory graft-versus-host disease. Leukemia. 2014;28:970-973. DOI:10.1038/leu.2014.41.

101. Lee HJ, Lee EG, Kang S, et al. Efficacy of microneedling plus human stem cell conditioned medium for skin reju-venation: a randomized, controlled, blinded split face study. Ann Dermatol. 2014;26:584-591.

102. Harn HJ, Huang MH, Huang CT, et al. Rejuvenation of aged pig facial skin by transplanting allogeneic granulo-cyte colony stimulating factor induced peripheral blood stem cells from a young pig. Cell Transplant. 2013;22: 755-765.

103. Kim WS, Park BS, Park SH, et al. Antiwrinkle effect of adipose-derived stem cell: activation of dermal fibroblast by secretory factors. J Dermatol Sci. 2009;53:96-102.

104. Shim JH, Park JY, Lee MG, et al. Human dermal stem/ progenitor cell-derived conditioned medium ameliorates ultraviolet a induced damage of normal human dermal fibroblasts. PLoS One 2013;e67604. DOI:10.1371/journal. pone.0067604.

105. Chen CC, Murray PJ, Jiang TX, et al. Regenerative hair waves in aging mice and extra-follicular modulators Follistatin, Dkk1 and Sfrp4. J Invest Dermatol. 2014;134:2086-2096.

106. Fukuoka H, Suga H. Hair regeneration treatment using adipose-derived stem cell conditioned medium: follow-up with trichograms. Eplasty. 2015;15:e10.

107. Ahmed MI, Alam M, Emelianov VU, et al. MicroRNA-214 controls skin and hair follicle development by modulat-ing the activity of the Wnt pathway. J Cell Biol. 2014;207:549-567.

108. Zhang GE, Grizzle WE. Exosomes: a novel pathway of local and distant intercellular communication that facil-itates the growth and metastasis of neoplastic lesions. Am J Pathol. 2014;184:28-41.

109. Harris DA, Patel SH, Gucek M, et al. Exosomes released from breast cancer carcinomas stimulate cell movement. PLoS One 2015;e0117495. DOI:10.1371/journal.pone.0117495.

111. Li J, Sherman-Baust CA, Tsai-Turton M, et al. Claudin containing exosomes in the peripheral circulation of women with ovarian cancer. BMC Cancer. 2009;9:244. DOI:10.1186/1471-2407-9-244.

118. Mittelbrunn M, Gutierrez-Vazquez C, Villarroya-Beltri C, et al. Unidirectional transfer of microRNA-loaded exo-somes from T cells to antigen-presenting cells. Nature Commun. 2011;2:282. DOI:10.1038/ncomms1285.

119. www.systembio.com [cited 2015 Apr 11].

120. Basu J, Ludlow JW. MSC sourced exosomes as therapeu-tic agents for wound healing and skin regeneration: from scaled production to functional regenerative out-comes in vitro and in vivo. International Society for Stem Cell Research Annual Meeting; 2015 Jun 24-27; Stockholm.

121. Basu J, Ludlow JW. Developmental engineering the kid-ney: leveraging principles of morphogenesis for renal regeneration. Birth Defects Res C Embryo Today. 2012;96:30-38.

122. Genheimer CW, Ilagan RM, Spencer T, et al. Molecular characterization of the regenerative response induced by intrarenal transplantation of selected renal cells in a rodent model of chronic kidney disease. Cells Tissues Organs. 2012;196:374-384.

123. www.fda.gov/BiologicsBloodVaccines/default.htm [cited2015 Apr 11].

A 99.9% Pure Exosome Intravenous Therapy Processed from Cultured Supernatant of Deciduous Dental Pulp Stem Cells

What are Exosomes?

Treatment Using Exosomes at Our Clinic