The establishment of a bank of stored clinical bone marrow stromal cell products

Abstract

BackgroundBone marrow stromal cells (BMSCs) are being used to treat a variety of conditions. For many applications a supply of cryopreserved products that can be used for acute therapy is needed. The establishment of a bank of BMSC products from healthy third party donors is described.MethodsThe recruitment of healthy subjects willing to donate marrow for BMSC production and the Good Manufacturing Practices (GMP) used for assessing potential donors, collecting marrow, culturing BMSCs and BMSC cryopreservation are described.ResultsSeventeen subjects were enrolled in our marrow collection protocol for BMSC production. Six of the 17 subjects were found to be ineligible during the donor screening process and one became ill and their donation was cancelled. Approximately 12 ml of marrow was aspirated from one posterior iliac crest of 10 donors; one donor donated twice. The BMSCs were initially cultured in T-75 flasks and then expanded for three passages in multilayer cell factories. The final BMSC product was packaged into units of 100 × 106 viable cells, cryopreserved and stored in a vapor phase liquid nitrogen tank under continuous monitoring. BMSC products meeting all lot release criteria were obtained from 8 of the 11 marrow collections. The rate of growth of the primary cultures was similar for all products except those generated from the two oldest donors. One lot did not meet the criteria for final release; its CD34 antigen expression was greater than the cut off set at 5%. The mean number of BMSC units obtained from each donor was 17 and ranged from 3 to 40.ConclusionsThe production of large numbers of BMSCs from bone marrow aspirates of healthy donors is feasible, but is limited by the high number of donors that did not meet eligibility criteria and products that did not meet lot release criteria.

Mesenchymal stromal cells from pooled mononuclear cells of multiple bone marrow donors as rescue therapy in pediatric severe steroid-refractory graft-versus-host disease: a multicenter survey

“…To our knowledge, there are few established MSC banks worldwide, and these MSC banks were generated by separately isolating, expanding, and freezing MSCs from up to 10 donors in FBS-containing media. [23][24][25][26] In the current study, we report for the first time the establishment of a serum-free and GMP-compliant MSC bank generated from pooled bone marrow mononuclear cells (BM-MNCs) of multiple donors as a novel strategy to circumvent donor-to-donor variability. Clinical-grade MSC endproducts (MEPs) derived from the MSC bank were thoroughly assessed for their proliferation, differentiation, and, in particular, for the allosuppressive potential in vitro.…”

Establishing a Bone Marrow Stromal Cell Transplant Program at the National Institutes of Health Clinical Center

“…17 Currently, we are using BMSCs from this repository to treat patients with acute GVHD, marrow failure, and tissue injury after hematopoietic stem cell transplantation and inflammatory bowel disease. This repository is needed because subjects with acute GVHD or inflammatory bowel disease require treatment prior to the *4 weeks required to isolate and expand BMSCs from a marrow aspirate.…”

“…The methods used to culture BMSC for the third party donor repository] were described by Sabatino et al 17 Marrow was aspirated from the posterior iliac crest of healthy subjects under an IRB approved protocol. A single cell suspension of marrow was prepared; without red blood cell lysis or density gradient separation.…”

Treating childhood traumatic brain injury with autologous stem cell therapy

“…The use of BMSCs does not encounter the same ethical dilemmas as ownership is clear and informed consent is given before donation. The NIH Bone Marrow Stromal Cell Transplantation Center (BMSC TC) was created in 2008 to create an infrastructure for the manufacture of clinical grade human BMSCs and to facilitate the use of ex vivo expanded BMSCs for the treatment of patients in the clinical setting [54]. …”