title: Functional maintenance of differentiated embryoid bodies in microfluidic systems: a platform for personalized medicine.
abstract: Hormone replacement therapies have become important for treating diseases such as premature ovarian failure or menopausal complications. The clinical use of bioidentical hormones might significantly reduce some of the potential risks reportedly associated with the use of synthetic hormones. In the present study, we demonstrate the utility and advantage of a microfluidic chip culture system to enhance the development of personalized, on-demand, treatment modules using embryoid bodies (EBs). Functional EBs cultured on microfluidic chips represent a platform for personalized, patient-specific treatment cassettes that can be cryopreserved until required for treatment. We assessed the viability, differentiation, and functionality of EBs cultured and cryopreserved in this system. During extended microfluidic culture, estradiol, progesterone, testosterone, and anti-müllerian hormone levels were measured, and the expression of differentiated steroidogenic cells was confirmed by immunocytochemistry assay for the ovarian tissue markers anti-müllerian hormone receptor type II, follicle-stimulating hormone receptor, and inhibin β-A and the estrogen biosynthesis enzyme aromatase. Our studies showed that under microfluidic conditions, differentiated steroidogenic EBs continued to secrete estradiol and progesterone at physiologically relevant concentrations (30-120 pg/ml and 150-450 pg/ml, respectively) for up to 21 days. Collectively, we have demonstrated for the first time the feasibility of using a microfluidic chip system with continuous flow for the differentiation and extended culture of functional steroidogenic stem cell-derived EBs, the differentiation of EBs into cells expressing ovarian antigens in a microfluidic system, and the ability to cryopreserve this system with restoration of growth and functionality on thawing. These results present a platform for the development of a new therapeutic system for personalized medicine.
authors: BAMM Labs, Canary Center at Stanford for Early Cancer Detection, Stanford University School of Medicine, Palo Alto, California, USA; BAMM Labs, Department of Medicine and Center for Infertility and Reproductive Surgery, Obstetrics Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
authors: Sinan
authors: S
authors: Guven
authors: BAMM Labs, Canary Center at Stanford for Early Cancer Detection, Stanford University School of Medicine, Palo Alto, California, USA; BAMM Labs, Department of Medicine and Center for Infertility and Reproductive Surgery, Obstetrics Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
authors: Jennifer S
authors: JS
authors: Lindsey
authors: BAMM Labs, Canary Center at Stanford for Early Cancer Detection, Stanford University School of Medicine, Palo Alto, California, USA; BAMM Labs, Department of Medicine and Center for Infertility and Reproductive Surgery, Obstetrics Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
authors: Ishwari
authors: I
authors: Poudel
authors: BAMM Labs, Canary Center at Stanford for Early Cancer Detection, Stanford University School of Medicine, Palo Alto, California, USA; BAMM Labs, Department of Medicine and Center for Infertility and Reproductive Surgery, Obstetrics Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
authors: Sireesha
authors: S
authors: Chinthala
authors: BAMM Labs, Canary Center at Stanford for Early Cancer Detection, Stanford University School of Medicine, Palo Alto, California, USA; BAMM Labs, Department of Medicine and Center for Infertility and Reproductive Surgery, Obstetrics Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
authors: Michael D
authors: MD
authors: Nickerson
authors: BAMM Labs, Canary Center at Stanford for Early Cancer Detection, Stanford University School of Medicine, Palo Alto, California, USA; BAMM Labs, Department of Medicine and Center for Infertility and Reproductive Surgery, Obstetrics Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
authors: Behzad
authors: B
authors: Gerami-Naini
authors: BAMM Labs, Canary Center at Stanford for Early Cancer Detection, Stanford University School of Medicine, Palo Alto, California, USA; BAMM Labs, Department of Medicine and Center for Infertility and Reproductive Surgery, Obstetrics Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
authors: Umut A
authors: UA
authors: Gurkan
authors: BAMM Labs, Canary Center at Stanford for Early Cancer Detection, Stanford University School of Medicine, Palo Alto, California, USA; BAMM Labs, Department of Medicine and Center for Infertility and Reproductive Surgery, Obstetrics Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA utkan@stanford.edu ranchan@partners.org.
authors: Raymond M
authors: RM
authors: Anchan
authors: BAMM Labs, Canary Center at Stanford for Early Cancer Detection, Stanford University School of Medicine, Palo Alto, California, USA; BAMM Labs, Department of Medicine and Center for Infertility and Reproductive Surgery, Obstetrics Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA utkan@stanford.edu ranchan@partners.org.
authors: Utkan
authors: U
authors: Demirci
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000378:metabolism; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D058732:Embryoid Bodies / Q000166:cytology / Q000378:metabolism; D006801:Humans; D051379:Mice; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods; D057285:Precision Medicine / Q000295:instrumentation / Q000379:methods
chemical_list: D000943:Antigens, Differentiation
keywords: Cryopreservation; Hormone secretion; Microenvironment; Microphysiological systems; Reproductive medicine; Stem cells