title: Manufacture of Clinical-Grade Human Clonal Mesenchymal Stem Cell Products from Single Colony Forming Unit-Derived Colonies Based on the Subfractionation Culturing Method.
abstract: Stem cell products derived from mesenchymal stem cells (MSCs) have been widely used in clinical trials, and a few products have been already commercialized. However, the therapeutic effects of clinical-grade MSCs are still controversial owing to mixed results from recent clinical trials. A potential solution to overcome this hurdle may be to use clonal stem cells as the starting cell material to increase the homogeneity of the final stem cell products. We have previously developed an alternative isolation and culture protocol for establishing a population of clonal MSCs (cMSCs) from single colony forming unit (CFU)-derived colonies. In this study, we established a good manufacturing practice (GMP)-compatible procedure for the clinical-grade production of human bone marrow-derived cMSCs based on the subfractionation culturing method. We optimized the culture procedures to expand and obtain a clonal population of final MSC products from single CFU-derived colonies in a GMP facility. The characterization results of the final cMSC products met our preset criteria. Animal toxicity tests were performed in a good laboratory practice facility, and showed no toxicity or tumor formation in vivo. These tests include single injection toxicity, multiple injection toxicity, biodistribution analysis, and tumorigenicity tests in vivo. No chromosomal abnormalities were detected by in situ karyotyping using oligo-fluorescence in situ hydridization (oligo-FISH), providing evidence of genetic stability of the clinical-grade cMSC products. The manufacture and quality control results indicated that our GMP methodology could produce sufficient clonal population of MSC products from a small amount of bone marrow aspirate to treat a number of patients.
authors.affiliation: 1 Translational Research Center, Inha University School of Medicine , Incheon, Republic of Korea.|2 Inha Research Institute for Medical Science, Inha University School of Medicine , Incheon, Republic of Korea.|3 SCM Lifescience Co., Ltd. , Incheon, Republic of Korea.
authors.forename: TacGhee
authors.initials: T
authors.lastname: Yi
authors.affiliation: 4 Drug Development Program, Department of Biomedical Science, Inha University School of Medicine , Incheon, Republic of Korea.
authors.forename: Si-na
authors.initials: SN
authors.lastname: Kim
authors.affiliation: 4 Drug Development Program, Department of Biomedical Science, Inha University School of Medicine , Incheon, Republic of Korea.
authors.forename: Hyun-Joo
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: 4 Drug Development Program, Department of Biomedical Science, Inha University School of Medicine , Incheon, Republic of Korea.
authors.forename: Junghee
authors.initials: J
authors.lastname: Kim
authors.affiliation: 3 SCM Lifescience Co., Ltd. , Incheon, Republic of Korea.
authors.forename: Yun-Kyoung
authors.initials: YK
authors.lastname: Cho
authors.affiliation: 1 Translational Research Center, Inha University School of Medicine , Incheon, Republic of Korea.|2 Inha Research Institute for Medical Science, Inha University School of Medicine , Incheon, Republic of Korea.
authors.forename: Dong-Hee
authors.initials: DH
authors.lastname: Shin
authors.affiliation: 1 Translational Research Center, Inha University School of Medicine , Incheon, Republic of Korea.
authors.forename: Sun-Ji
authors.initials: SJ
authors.lastname: Tak
authors.affiliation: 3 SCM Lifescience Co., Ltd. , Incheon, Republic of Korea.
authors.forename: Sun-Hwa
authors.initials: SH
authors.lastname: Moon
authors.affiliation: 3 SCM Lifescience Co., Ltd. , Incheon, Republic of Korea.
authors.forename: Ji-Eun
authors.initials: JE
authors.lastname: Kang
authors.affiliation: 3 SCM Lifescience Co., Ltd. , Incheon, Republic of Korea.
authors.forename: In-Mi
authors.initials: IM
authors.lastname: Ji
authors.affiliation: 3 SCM Lifescience Co., Ltd. , Incheon, Republic of Korea.
authors.forename: Huyn-Ja
authors.initials: HJ
authors.lastname: Lim
authors.affiliation: 5 Department of Pathology, Seoul National University School of Medicine , Seoul, Republic of Korea.
authors.forename: Dong-Soon
authors.initials: DS
authors.lastname: Lee
authors.affiliation: 1 Translational Research Center, Inha University School of Medicine , Incheon, Republic of Korea.
authors.forename: Myung-Shin
authors.initials: MS
authors.lastname: Jeon
authors.affiliation: 1 Translational Research Center, Inha University School of Medicine , Incheon, Republic of Korea.|3 SCM Lifescience Co., Ltd. , Incheon, Republic of Korea.
authors.forename: Sun U
authors.initials: SU
authors.lastname: Song
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*