Stem cell transplant Flashcards
BM collection has lower ______ content than PBSC and do not require mobilization
T cell
Target cell dose for BM collection
2 x 10^8 TNC/kg
Target cell dose for PBSC
> 5 X 10^6 CD34/kg
HLA matching appears to demonstrate survival benefit at this stage of disease
early and intermediate
risk of GVHD associated with HLA-DPB1 mismatching was influenced by the HLA-DPB1 rs9277534 expression marker, with _____ subtype being associated with greater risk than ______ subtype
G;A
Melting Temperature equation
Tm= 4(G+C) + 2 (A+T)
Temperature and salt conditions favored for high stringency
High and low
PCR: temperature at which denaturation occurs
94-96C
PCR: temperature at which primers anneal
50-65C
PCR: temperature at which extention occurs
72C
Too much Mg can decrease _______
fidelity
Variable EDTA and Citrate in sample can effect amount of _____
MG
Mg required for ______ activation
Taq polymerase
KCl important in _______
primer annealing
DMSO can be added and temperatures lowered when ____ content is high; however this can negatively affect_____
GC; Taq
SSP requires ______ specificity
3’end
Ideal primer length
~20 nucleotides
Ideal primer GC content; however avoid ______
50%; GC runs
Probe design: best specificity lies in ______ at the _____ of the probe
polymorphism; center
Early CD3 engraftment may identify patients at risk of _____
GVHD
How many cells do you need for 1 ng of template?
150
How many cells do you need for 10 ng of template?
1500
How many cell do you need for 100 ng of template
150,000
Minimum dose of single cord transplant
3 x 10^7 TNC/kg
http://hla.alleles.org
Nomenclature
http://igdawg.org/cwd.html
Common and well documented alleles
http://www.ebi.ac.uk/ipd/imgt/hla/
Allele history, alignments, serologic links
https://bioinformatics.bethematchcli nical.org/
Allele/haplotype frequencies in US, haplotype tools, multi-allele codes, link to NMDP search strategy advice, CT policies
http://www.pypop.org/popdata/
World-wide allele distribution maps
http://www.bmdw.org/
World-wide database HLA phenotypes of volunteer donors, cbu for transplant centers/search coordinators; requires authorization to access
National registries / cord blood banks; USA–
https://bethematchclinical.org/
Preliminary & formal unrelated donor/cbu search (physician (prelim only), transplant center)
https://www.mh-hannover.de/institute/transfusion/histocheck/
visualizing ribbon structure of HLA molecule and interrogation of peptide groove
______ are rules mandated by the government
regulations
______ are rules established by accrediting agency
standards
STR: Mixture of donor and host
Mixed chimerism
STR: One or more lineages complete donor others complete host
Split chimerism
STR: Mixed chimerism in a subset (e.g., T cells)
Subset chimerism
STR: <1% donor chimerism
microchimerim
STRs are this long and are known as _______
2-8 basepairs(some variability); microsatellites
VNTR are this long and are known as _______
> 8 basepairs, usually much larger; minisatellites
STR: Repeats that are short sequences of the same length
Simple
STR: 2 or more adjacent repeats of same length
Compound
STR: Variable length and composition repeats
Complex
